Principles & Protocols

Protocols - Grid Preparation

• Solid Carbon Films
• Triafol Holey Films
• Formvar Holey Films
• Carbon sandwich
• Fiducials for tomography

Principles

Grid selection

Shiny vs. Dull side

Mesh Size (lines per inch)

• 400 mesh - recommended for support films. Protein imaging applications, for example.
• 200 mesh - A good compromise for tomography.
• Slotted grids - For tomography and serial sections. Need to be coated with a plastic film.

Material

Common

• Copper - Most common - inexpensive, conductive, beam stable. Relatively thin grid bars.
• Copper/Palladium - Palladium adds rigidity to the grid, but makes it more expensive.
• Copper/Rhodium - non-tarnishing, reduction in bar thickness. Recognizable sides.

Less common

• Gold or Platinum - non-reactive, expensive, beam stable. Easily bent (especially gold)
• Molybdenum - Expensive, Coefficient of contraction similar to carbon. Quite rigid.
• Nickel - Used mainly for silver enhancement.
• Beryllium - Minimize background radiation. Mainly used for material science.
• Titanium - Extra stiff.

Mesh types

• Square Mesh. These are by far the most common. There are many brands with different peculiarities each.
• Finder Grids - These are particularly useful when one needs to go back to collect data from a known area.
• Hexagonal Mesh - Used for tomography more than anything, but can be used for the same applications as the square grids.
• Slot Grids - For tomography. More fragile than others since there is less support

Support films

With CarbonCoating

• Formvar and Carbon - A formvar film is made on a glass slide. Then the film is floated in water and grids placed on it. The grids are then picked up with paper from the water surface and carbon is evaporated on top of the formvar.
• Carbon only - In this case, carbon is evaporated on a freshly cleaved mica sheet. The carbon film is then floated off of the mica and lowered onto submerged grids placed on a piece of filter paper.
• Nitrocellulose and Carbon - Similar to the formvar-carbon film. A drop of nitrocellulose is placed on a crystallization dish, forming thus a thin plastic film. Grids are placed on the surface, picked up with paper and coated with carbon.

• Carbon with holes - These films are used commonly for frozen hydrated samples. There are several ways to prepare them, or they can be purchased from vendors (Quantifoil, C-flat). Commercially available grids have regular holes.
  • Quantifoil - commercially available. Carbon is ~200 A thick, sturdy, but generally not suitable for imaging. A thin coating of carbon can be floated on these grids if the sample always settles on carbon and not the holes.
  • C-flat - commercially available. Thinner than Quantifoil, but more fragile. May be able to image over this carbon
  • Lacey Carbon - Home-made plastic films which are carbon coated, then plastic is dissolved away. These grids can be used to support tissue sections, or can serve as support for a very thin carbon film and used also for single particle analysis.

Uncommon coatings

• • Beryllium
  • Silicone monoxide - low background contrast, stable under beam, more hydrophilic than carbon

• Set ALLOWTOPICCHANGE = CemfacGroup

-- KdDerr - 28 Aug 2007

Principles & Protocols

Protocols - Grid Preparation

• Solid Carbon Films
• Triafol Holey Films
• Formvar Holey Films
• Carbon sandwich
• Fiducials for tomography
• Nitrocellulose Film

Principles

Grid selection

Shiny vs. Dull side

Mesh Size (lines per inch)

• 400 mesh - recommended for support films. Protein imaging applications, for example.
• 200 mesh - A good compromise for tomography.
• Slotted grids - For tomography and serial sections. Need to be coated with a plastic film.

Material

Common

• Copper - Most common - inexpensive, conductive, beam stable. Relatively thin grid bars.
• Copper/Palladium - Palladium adds rigidity to the grid, but makes it more expensive.
• Copper/Rhodium - non-tarnishing, reduction in bar thickness. Recognizable sides.

Less common

• Gold or Platinum - non-reactive, expensive, beam stable. Easily bent (especially gold)
• Molybdenum - Expensive, Coefficient of contraction similar to carbon. Quite rigid.
• Nickel - Used mainly for silver enhancement.
• Beryllium - Minimize background radiation. Mainly used for material science.
• Titanium - Extra stiff.

Mesh types

• Square Mesh. These are by far the most common. There are many brands with different peculiarities each.
• Finder Grids - These are particularly useful when one needs to go back to collect data from a known area.
• Hexagonal Mesh - Used for tomography more than anything, but can be used for the same applications as the square grids.
• Slot Grids - For tomography. More fragile than others since there is less support

Support films

With CarbonCoating

• Formvar and Carbon - A formvar film is made on a glass slide. Then the film is floated in water and grids placed on it. The grids are then picked up with paper from the water surface and carbon is evaporated on top of the formvar.
• Carbon only - In this case, carbon is evaporated on a freshly cleaved mica sheet. The carbon film is then floated off of the mica and lowered onto submerged grids placed on a piece of filter paper.
• Nitrocellulose and Carbon - Similar to the formvar-carbon film. A drop of nitrocellulose is placed on a crystallization dish, forming thus a thin plastic film. Grids are placed on the surface, picked up with paper and coated with carbon.

• Carbon with holes - These films are used commonly for frozen hydrated samples. There are several ways to prepare them, or they can be purchased from vendors (Quantifoil, C-flat). Commercially available grids have regular holes.
  • Quantifoil - commercially available. Carbon is ~200 A thick, sturdy, but generally not suitable for imaging. A thin coating of carbon can be floated on these grids if the sample always settles on carbon and not the holes.
  • C-flat - commercially available. Thinner than Quantifoil, but more fragile. May be able to image over this carbon
  • Lacey Carbon - Home-made plastic films which are carbon coated, then plastic is dissolved away. These grids can be used to support tissue sections, or can serve as support for a very thin carbon film and used also for single particle analysis.

Uncommon coatings

• • Beryllium
  • Silicone monoxide - low background contrast, stable under beam, more hydrophilic than carbon

• Set ALLOWTOPICCHANGE = CemfacGroup

-- KdDerr - 28 Aug 2007

Principles & Protocols

Protocols - Grid Preparation

• Solid Carbon Films
• Triafol Holey Films
• Formvar Holey Films
• Carbon sandwich
• Fiducials for tomography

Principles

Grid selection

Shiny vs. Dull side

Mesh Size (lines per inch)

• 400 mesh - recommended for support films. Protein imaging applications, for example.
• 200 mesh - A good compromise for tomography.
• Slotted grids - For tomography and serial sections. Need to be coated with a plastic film.

Material

Common

• Copper - Most common - inexpensive, conductive, beam stable. Relatively thin grid bars.
• Copper/Palladium - Palladium adds rigidity to the grid, but makes it more expensive.
• Copper/Rhodium - non-tarnishing, reduction in bar thickness. Recognizable sides.

Less common

• Gold or Platinum - non-reactive, expensive, beam stable. Easily bent (especially gold)
• Molybdenum - Expensive, Coefficient of contraction similar to carbon. Quite rigid.
• Nickel - Used mainly for silver enhancement.
• Beryllium - Minimize background radiation. Mainly used for material science.
• Titanium - Extra stiff.

Mesh types

• Square Mesh. These are by far the most common. There are many brands with different peculiarities each.
• Finder Grids - These are particularly useful when one needs to go back to collect data from a known area.
• Hexagonal Mesh - Used for tomography more than anything, but can be used for the same applications as the square grids.
• Slot Grids - For tomography. More fragile than others since there is less support

Support films

With CarbonCoating

• Formvar and Carbon - A formvar film is made on a glass slide. Then the film is floated in water and grids placed on it. The grids are then picked up with paper from the water surface and carbon is evaporated on top of the formvar.
• Carbon only - In this case, carbon is evaporated on a freshly cleaved mica sheet. The carbon film is then floated off of the mica and lowered onto submerged grids placed on a piece of filter paper.
• Nitrocellulose and Carbon - Similar to the formvar-carbon film. A drop of nitrocellulose is placed on a crystallization dish, forming thus a thin plastic film. Grids are placed on the surface, picked up with paper and coated with carbon.

• Carbon with holes - These films are used commonly for frozen hydrated samples. There are several ways to prepare them, or they can be purchased from vendors (Quantifoil, C-flat). Commercially available grids have regular holes.
  • Quantifoil - commercially available. Carbon is ~200 A thick, sturdy, but generally not suitable for imaging. A thin coating of carbon can be floated on these grids if the sample always settles on carbon and not the holes.
  • C-flat - commercially available. Thinner than Quantifoil, but more fragile. May be able to image over this carbon
  • Lacey Carbon - Home-made plastic films which are carbon coated, then plastic is dissolved away. These grids can be used to support tissue sections, or can serve as support for a very thin carbon film and used also for single particle analysis.

Uncommon coatings

• • Beryllium
  • Silicone monoxide - low background contrast, stable under beam, more hydrophilic than carbon

• Set ALLOWTOPICCHANGE = CemfacGroup

-- KdDerr - 28 Aug 2007

Principles & Protocols

Protocols - Grid Preparation

• Solid Carbon Films
• Triafol Holey Films
• Formvar Holey Films
• Carbon sandwich
• Fiducials for tomography

Principles

Grid selection

Shiny vs. Dull side

Mesh Size (lines per inch)

• 400 mesh - recommended for support films. Protein imaging applications, for example.
• 200 mesh - A good compromise for tomography.
• Slotted grids - For tomography and serial sections. Need to be coated with a plastic film.

Material

Common

• Copper - Most common - inexpensive, conductive, beam stable. Relatively thin grid bars.
• Copper/Palladium - Palladium adds rigidity to the grid, but makes it more expensive.
• Copper/Rhodium - non-tarnishing, reduction in bar thickness. Recognizable sides.

Less common

• Gold or Platinum - non-reactive, expensive, beam stable. Easily bent (especially gold)
• Molybdenum - Expensive, Coefficient of contraction similar to carbon. Quite rigid.
• Nickel - Used mainly for silver enhancement.
• Beryllium - Minimize background radiation. Mainly used for material science.
• Titanium - Extra stiff.

Mesh types

• Square Mesh. These are by far the most common. There are many brands with different peculiarities each.
• Finder Grids - These are particularly useful when one needs to go back to collect data from a known area.
• Hexagonal Mesh - Used for tomography more than anything, but can be used for the same applications as the square grids.
• Slot Grids - For tomography. More fragile than others since there is less support

Support films

With CarbonCoating

• Formvar and Carbon - A formvar film is made on a glass slide. Then the film is floated in water and grids placed on it. The grids are then picked up with paper from the water surface and carbon is evaporated on top of the formvar.
• Carbon only - In this case, carbon is evaporated on a freshly cleaved mica sheet. The carbon film is then floated off of the mica and lowered onto submerged grids placed on a piece of filter paper.
• Nitrocellulose and Carbon - Similar to the formvar-carbon film. A drop of nitrocellulose is placed on a crystallization dish, forming thus a thin plastic film. Grids are placed on the surface, picked up with paper and coated with carbon.

• Carbon with holes - These films are used commonly for frozen hydrated samples. There are several ways to prepare them, or they can be purchased from vendors (Quantifoil, C-flat). Commercially available grids have regular holes.
  • Quantifoil - commercially available. Carbon is ~200 A thick, sturdy, but generally not suitable for imaging. A thin coating of carbon can be floated on these grids if the sample always settles on carbon and not the holes.
  • C-flat - commercially available. Thinner than Quantifoil, but more fragile. May be able to image over this carbon
  • Lacey Carbon - Home-made plastic films which are carbon coated, then plastic is dissolved away. These grids can be used to support tissue sections, or can serve as support for a very thin carbon film and used also for single particle analysis.

Uncommon coatings

• • Beryllium
  • Silicone monoxide - low background contrast, stable under beam, more hydrophilic than carbon

• Set ALLOWTOPICCHANGE = CemfacGroup

-- KdDerr - 28 Aug 2007

Protocols - Grid Preparation

• Solid Carbon Films
• Triafol Holey Films
• Formvar Holey Films
• Carbon sandwich
• Fiducials for tomography
• PlasticCoatedGrid

Principles

Grid selection

Shiny vs. Dull side

Mesh Size (lines per inch)

• 400 mesh - recommended for support films. Protein imaging applications, for example.
• 200 mesh - A good compromise for tomography.
• Slotted grids - For tomography and serial sections. Need to be coated with a plastic film.

Material

Common

• Copper - Most common - inexpensive, conductive, beam stable. Relatively thin grid bars.
• Copper/Palladium - Palladium adds rigidity to the grid, but makes it more expensive.
• Copper/Rhodium - non-tarnishing, reduction in bar thickness. Recognizable sides.

Less common

• Gold or Platinum - non-reactive, expensive, beam stable. Easily bent (especially gold)
• Molybdenum - Expensive, Coefficient of contraction similar to carbon. Quite rigid.
• Nickel - Used mainly for silver enhancement.
• Beryllium - Minimize background radiation. Mainly used for material science.
• Titanium - Extra stiff.

Mesh types

• Square Mesh. These are by far the most common. There are many brands with different peculiarities each.
• Finder Grids - These are particularly useful when one needs to go back to collect data from a known area.
• Hexagonal Mesh - Used for tomography more than anything, but can be used for the same applications as the square grids.
• Slot Grids - For tomography. More fragile than others since there is less support

Support films

With CarbonCoating

• Formvar and Carbon - A formvar film is made on a glass slide. Then the film is floated in water and grids placed on it. The grids are then picked up with paper from the water surface and carbon is evaporated on top of the formvar.
• Carbon only - In this case, carbon is evaporated on a freshly cleaved mica sheet. The carbon film is then floated off of the mica and lowered onto submerged grids placed on a piece of filter paper.
• Nitrocellulose and Carbon - Similar to the formvar-carbon film. A drop of nitrocellulose is placed on a crystallization dish, forming thus a thin plastic film. Grids are placed on the surface, picked up with paper and coated with carbon.

• Carbon with holes - These films are used commonly for frozen hydrated samples. There are several ways to prepare them, or they can be purchased from vendors (Quantifoil, C-flat). Commercially available grids have regular holes.
  • Quantifoil - commercially available. Carbon is ~200 A thick, sturdy, but generally not suitable for imaging. A thin coating of carbon can be floated on these grids if the sample always settles on carbon and not the holes.
  • C-flat - commercially available. Thinner than Quantifoil, but more fragile. May be able to image over this carbon
  • Lacey Carbon - Home-made plastic films which are carbon coated, then plastic is dissolved away. These grids can be used to support tissue sections, or can serve as support for a very thin carbon film and used also for single particle analysis.

Uncommon coatings

• • Beryllium
  • Silicone monoxide - low background contrast, stable under beam, more hydrophilic than carbon

• Set ALLOWTOPICCHANGE = CemfacGroup

-- KdDerr - 28 Aug 2007

EM Grids: Principles & Protocols

• Solid Carbon Films
• Triafol Holey Films
• Formvar Holey Films
• Carbon sandwich
• Fiducials for tomography
• PlasticCoatedGrid

Principles

Grid selection

Shiny vs. Dull side

Mesh Size (lines per inch)

• 400 mesh - recommended for support films. Protein imaging applications, for example.
• 200 mesh - A good compromise for tomography.
• Slotted grids - For tomography and serial sections. Need to be coated with a plastic film.

Material

Common

• Copper - Most common - inexpensive, conductive, beam stable. Relatively thin grid bars.
• Copper/Palladium - Palladium adds rigidity to the grid, but makes it more expensive.
• Copper/Rhodium - non-tarnishing, reduction in bar thickness. Recognizable sides.

Less common

• Gold or Platinum - non-reactive, expensive, beam stable. Easily bent (especially gold)
• Molybdenum - Expensive, Coefficient of contraction similar to carbon. Quite rigid.
• Nickel - Used mainly for silver enhancement.
• Beryllium - Minimize background radiation. Mainly used for material science.
• Titanium - Extra stiff.

Mesh types

• Square Mesh. These are by far the most common. There are many brands with different peculiarities each.
• Finder Grids - These are particularly useful when one needs to go back to collect data from a known area.
• Hexagonal Mesh - Used for tomography more than anything, but can be used for the same applications as the square grids.
• Slot Grids - For tomography. More fragile than others since there is less support

Support films

With CarbonCoating

• Formvar and Carbon - A formvar film is made on a glass slide. Then the film is floated in water and grids placed on it. The grids are then picked up with paper from the water surface and carbon is evaporated on top of the formvar.
• Carbon only - In this case, carbon is evaporated on a freshly cleaved mica sheet. The carbon film is then floated off of the mica and lowered onto submerged grids placed on a piece of filter paper.
• Nitrocellulose and Carbon - Similar to the formvar-carbon film. A drop of nitrocellulose is placed on a crystallization dish, forming thus a thin plastic film. Grids are placed on the surface, picked up with paper and coated with carbon.

• Carbon with holes - These films are used commonly for frozen hydrated samples. There are several ways to prepare them, or they can be purchased from vendors (Quantifoil, C-flat). Commercially available grids have regular holes.
  • Quantifoil - commercially available. Carbon is ~200 A thick, sturdy, but generally not suitable for imaging. A thin coating of carbon can be floated on these grids if the sample always settles on carbon and not the holes.
  • C-flat - commercially available. Thinner than Quantifoil, but more fragile. May be able to image over this carbon
  • Lacey Carbon - Home-made plastic films which are carbon coated, then plastic is dissolved away. These grids can be used to support tissue sections, or can serve as support for a very thin carbon film and used also for single particle analysis.

Uncommon coatings

• • Beryllium
  • Silicone monoxide - low background contrast, stable under beam, more hydrophilic than carbon

• Set ALLOWTOPICCHANGE = CemfacGroup

-- KdDerr - 28 Aug 2007

EM Grids: Principles & Protocols

Protocols

Grid Preparation

• Solid Carbon Films
• Triafol Holey Films
• Formvar Holey Films
• Carbon sandwich
• Fiducials for tomography

Principles

Grid selection

Shiny vs. Dull side

Mesh Size (lines per inch)

• 400 mesh - recommended for support films. Protein imaging applications, for example.
• 200 mesh - A good compromise for tomography.
• Slotted grids - For tomography and serial sections. Need to be coated with a plastic film.

Material

Common

• Copper - Most common - inexpensive, conductive, beam stable. Relatively thin grid bars.
• Copper/Palladium - Palladium adds rigidity to the grid, but makes it more expensive.
• Copper/Rhodium - non-tarnishing, reduction in bar thickness. Recognizable sides.

Less common

• Gold or Platinum - non-reactive, expensive, beam stable. Easily bent (especially gold)
• Molybdenum - Expensive, Coefficient of contraction similar to carbon. Quite rigid.
• Nickel - Used mainly for silver enhancement.
• Beryllium - Minimize background radiation. Mainly used for material science.
• Titanium - Extra stiff.

Mesh types

• Square Mesh. These are by far the most common. There are many brands with different peculiarities each.
• Finder Grids - These are particularly useful when one needs to go back to collect data from a known area.
• Hexagonal Mesh - Used for tomography more than anything, but can be used for the same applications as the square grids.
• Slot Grids - For tomography. More fragile than others since there is less support

Support films

With CarbonCoating

• Formvar and Carbon - A formvar film is made on a glass slide. Then the film is floated in water and grids placed on it. The grids are then picked up with paper from the water surface and carbon is evaporated on top of the formvar.
• Carbon only - In this case, carbon is evaporated on a freshly cleaved mica sheet. The carbon film is then floated off of the mica and lowered onto submerged grids placed on a piece of filter paper.
• Nitrocellulose and Carbon - Similar to the formvar-carbon film. A drop of nitrocellulose is placed on a crystallization dish, forming thus a thin plastic film. Grids are placed on the surface, picked up with paper and coated with carbon.

• Carbon with holes - These films are used commonly for frozen hydrated samples. There are several ways to prepare them, or they can be purchased from vendors (Quantifoil, C-flat). Commercially available grids have regular holes.
  • Quantifoil - commercially available. Carbon is ~200 A thick, sturdy, but generally not suitable for imaging. A thin coating of carbon can be floated on these grids if the sample always settles on carbon and not the holes.
  • C-flat - commercially available. Thinner than Quantifoil, but more fragile. May be able to image over this carbon
  • Lacey Carbon - Home-made plastic films which are carbon coated, then plastic is dissolved away. These grids can be used to support tissue sections, or can serve as support for a very thin carbon film and used also for single particle analysis.

Uncommon coatings

• • Beryllium
  • Silicone monoxide - low background contrast, stable under beam, more hydrophilic than carbon

• Set ALLOWTOPICCHANGE = CemfacGroup

-- KdDerr - 28 Aug 2007

EM Grids: Principles & Protocols

Protocols

Grid Preparation

• Solid Carbon Films
• Triafol Holey Films
• Formvar Holey Films
• Carbon sandwich
• Fiducials for tomography

Grid selection

Shiny vs. Dull side

Mesh Size (lines per inch)

• 400 mesh - recommended for support films. Protein imaging applications, for example.
• 200 mesh - A good compromise for tomography.
• Slotted grids - For tomography and serial sections. Need to be coated with a plastic film.

Material

Common

• Copper - Most common - inexpensive, conductive, beam stable. Relatively thin grid bars.
• Copper/Palladium - Palladium adds rigidity to the grid, but makes it more expensive.
• Copper/Rhodium - non-tarnishing, reduction in bar thickness. Recognizable sides.

Less common

• Gold or Platinum - non-reactive, expensive, beam stable. Easily bent (especially gold)
• Molybdenum - Expensive, Coefficient of contraction similar to carbon. Quite rigid.
• Nickel - Used mainly for silver enhancement.
• Beryllium - Minimize background radiation. Mainly used for material science.
• Titanium - Extra stiff.

Mesh types

• Square Mesh. These are by far the most common. There are many brands with different peculiarities each.
• Finder Grids - These are particularly useful when one needs to go back to collect data from a known area.
• Hexagonal Mesh - Used for tomography more than anything, but can be used for the same applications as the square grids.
• Slot Grids - For tomography. More fragile than others since there is less support

Support films

With CarbonCoating

• Formvar and Carbon - A formvar film is made on a glass slide. Then the film is floated in water and grids placed on it. The grids are then picked up with paper from the water surface and carbon is evaporated on top of the formvar.
• Carbon only - In this case, carbon is evaporated on a freshly cleaved mica sheet. The carbon film is then floated off of the mica and lowered onto submerged grids placed on a piece of filter paper.
• Nitrocellulose and Carbon - Similar to the formvar-carbon film. A drop of nitrocellulose is placed on a crystallization dish, forming thus a thin plastic film. Grids are placed on the surface, picked up with paper and coated with carbon.

• Carbon with holes - These films are used commonly for frozen hydrated samples. There are several ways to prepare them, or they can be purchased from vendors (Quantifoil, C-flat). Commercially available grids have regular holes.
  • Quantifoil - commercially available. Carbon is ~200 A thick, sturdy, but generally not suitable for imaging. A thin coating of carbon can be floated on these grids if the sample always settles on carbon and not the holes.
  • C-flat - commercially available. Thinner than Quantifoil, but more fragile. May be able to image over this carbon
  • Lacey Carbon - Home-made plastic films which are carbon coated, then plastic is dissolved away. These grids can be used to support tissue sections, or can serve as support for a very thin carbon film and used also for single particle analysis.

Uncommon coatings

• • Beryllium
  • Silicone monoxide - low background contrast, stable under beam, more hydrophilic than carbon

• Set ALLOWTOPICCHANGE = CemfacGroup

-- KdDerr - 28 Aug 2007

EM Grids: Principles & Protocols

Principles

Protocols

Grid Preparation

• Solid Carbon Films
• Triafol Holey Films
• Formvar Holey Films
• Carbon sandwich
• Fiducials for tomography

Grid selection

Shiny vs. Dull side

Mesh Size (lines per inch)

• 400 mesh - recommended for support films. Protein imaging applications, for example.
• 200 mesh - A good compromise for tomography.
• Slotted grids - For tomography and serial sections. Need to be coated with a plastic film.

Material

Common

• Copper - Most common - inexpensive, conductive, beam stable. Relatively thin grid bars.
• Copper/Palladium - Palladium adds rigidity to the grid, but makes it more expensive.
• Copper/Rhodium - non-tarnishing, reduction in bar thickness. Recognizable sides.

Less common

• Gold or Platinum - non-reactive, expensive, beam stable. Easily bent (especially gold)
• Molybdenum - Expensive, Coefficient of contraction similar to carbon. Quite rigid.
• Nickel - Used mainly for silver enhancement.
• Beryllium - Minimize background radiation. Mainly used for material science.
• Titanium - Extra stiff.

Mesh types

• Square Mesh. These are by far the most common. There are many brands with different peculiarities each.
• Finder Grids - These are particularly useful when one needs to go back to collect data from a known area.
• Hexagonal Mesh - Used for tomography more than anything, but can be used for the same applications as the square grids.
• Slot Grids - For tomography. More fragile than others since there is less support

Support films

With CarbonCoating

• Formvar and Carbon - A formvar film is made on a glass slide. Then the film is floated in water and grids placed on it. The grids are then picked up with paper from the water surface and carbon is evaporated on top of the formvar.
• Carbon only - In this case, carbon is evaporated on a freshly cleaved mica sheet. The carbon film is then floated off of the mica and lowered onto submerged grids placed on a piece of filter paper.
• Nitrocellulose and Carbon - Similar to the formvar-carbon film. A drop of nitrocellulose is placed on a crystallization dish, forming thus a thin plastic film. Grids are placed on the surface, picked up with paper and coated with carbon.

• Carbon with holes - These films are used commonly for frozen hydrated samples. There are several ways to prepare them, or they can be purchased from vendors (Quantifoil, C-flat). Commercially available grids have regular holes.
  • Quantifoil - commercially available. Carbon is ~200 A thick, sturdy, but generally not suitable for imaging. A thin coating of carbon can be floated on these grids if the sample always settles on carbon and not the holes.
  • C-flat - commercially available. Thinner than Quantifoil, but more fragile. May be able to image over this carbon
  • Lacey Carbon - Home-made plastic films which are carbon coated, then plastic is dissolved away. These grids can be used to support tissue sections, or can serve as support for a very thin carbon film and used also for single particle analysis.

Uncommon coatings

• • Beryllium
  • Silicone monoxide - low background contrast, stable under beam, more hydrophilic than carbon

• Set ALLOWTOPICCHANGE = CemfacGroup

-- KdDerr - 28 Aug 2007

EM Grids: Principles & Protocols

Principles

Protocols

Grid Preparation

• Solid Carbon Films
• Triafol Holey Films

• Carbon sandwich
• Fiducials for tomography

Grid selection

Shiny vs. Dull side

Mesh Size (lines per inch)

• 400 mesh - recommended for support films. Protein imaging applications, for example.
• 200 mesh - A good compromise for tomography.
• Slotted grids - For tomography and serial sections. Need to be coated with a plastic film.

Material

Common

• Copper - Most common - inexpensive, conductive, beam stable. Relatively thin grid bars.
• Copper/Palladium - Palladium adds rigidity to the grid, but makes it more expensive.
• Copper/Rhodium - non-tarnishing, reduction in bar thickness. Recognizable sides.

Less common

• Gold or Platinum - non-reactive, expensive, beam stable. Easily bent (especially gold)
• Molybdenum - Expensive, Coefficient of contraction similar to carbon. Quite rigid.
• Nickel - Used mainly for silver enhancement.
• Beryllium - Minimize background radiation. Mainly used for material science.
• Titanium - Extra stiff.

Mesh types

• Square Mesh. These are by far the most common. There are many brands with different peculiarities each.
• Finder Grids - These are particularly useful when one needs to go back to collect data from a known area.
• Hexagonal Mesh - Used for tomography more than anything, but can be used for the same applications as the square grids.
• Slot Grids - For tomography. More fragile than others since there is less support

Support films

With CarbonCoating

• Formvar and Carbon - A formvar film is made on a glass slide. Then the film is floated in water and grids placed on it. The grids are then picked up with paper from the water surface and carbon is evaporated on top of the formvar.
• Carbon only - In this case, carbon is evaporated on a freshly cleaved mica sheet. The carbon film is then floated off of the mica and lowered onto submerged grids placed on a piece of filter paper.
• Nitrocellulose and Carbon - Similar to the formvar-carbon film. A drop of nitrocellulose is placed on a crystallization dish, forming thus a thin plastic film. Grids are placed on the surface, picked up with paper and coated with carbon.

• Carbon with holes - These films are used commonly for frozen hydrated samples. There are several ways to prepare them, or they can be purchased from vendors (Quantifoil, C-flat). Commercially available grids have regular holes.
  • Quantifoil - commercially available. Carbon is ~200 A thick, sturdy, but generally not suitable for imaging. A thin coating of carbon can be floated on these grids if the sample always settles on carbon and not the holes.
  • C-flat - commercially available. Thinner than Quantifoil, but more fragile. May be able to image over this carbon
  • Lacey Carbon - Home-made plastic films which are carbon coated, then plastic is dissolved away. These grids can be used to support tissue sections, or can serve as support for a very thin carbon film and used also for single particle analysis.

Uncommon coatings

• • Beryllium
  • Silicone monoxide - low background contrast, stable under beam, more hydrophilic than carbon

• Set ALLOWTOPICCHANGE = CemfacGroup

-- KdDerr - 28 Aug 2007

EM Grids: Principles & Protocols

Principles

Protocols

Grid Preparation

• Solid Carbon Films
• Triafol Holey Films
• Claudio Holey Films
• Carbon sandwich
• Fiducials for tomography

Grid selection

Shiny vs. Dull side

Mesh Size (lines per inch)

• 400 mesh - recommended for support films. Protein imaging applications, for example.
• 200 mesh - A good compromise for tomography.
• Slotted grids - For tomography and serial sections. Need to be coated with a plastic film.

Material

Common

• Copper - Most common - inexpensive, conductive, beam stable. Relatively thin grid bars.
• Copper/Palladium - Palladium adds rigidity to the grid, but makes it more expensive.
• Copper/Rhodium - non-tarnishing, reduction in bar thickness. Recognizable sides.

Less common

• Gold or Platinum - non-reactive, expensive, beam stable. Easily bent (especially gold)
• Molybdenum - Expensive, Coefficient of contraction similar to carbon. Quite rigid.
• Nickel - Used mainly for silver enhancement.
• Beryllium - Minimize background radiation. Mainly used for material science.
• Titanium - Extra stiff.

Mesh types

• Square Mesh. These are by far the most common. There are many brands with different peculiarities each.
• Finder Grids - These are particularly useful when one needs to go back to collect data from a known area.
• Hexagonal Mesh - Used for tomography more than anything, but can be used for the same applications as the square grids.
• Slot Grids - For tomography. More fragile than others since there is less support

Support films

With CarbonCoating

• Formvar and Carbon - A formvar film is made on a glass slide. Then the film is floated in water and grids placed on it. The grids are then picked up with paper from the water surface and carbon is evaporated on top of the formvar.
• Carbon only - In this case, carbon is evaporated on a freshly cleaved mica sheet. The carbon film is then floated off of the mica and lowered onto submerged grids placed on a piece of filter paper.
• Nitrocellulose and Carbon - Similar to the formvar-carbon film. A drop of nitrocellulose is placed on a crystallization dish, forming thus a thin plastic film. Grids are placed on the surface, picked up with paper and coated with carbon.

• Carbon with holes - These films are used commonly for frozen hydrated samples. There are several ways to prepare them, or they can be purchased from vendors (Quantifoil, C-flat). Commercially available grids have regular holes.
  • Quantifoil - commercially available. Carbon is ~200 A thick, sturdy, but generally not suitable for imaging. A thin coating of carbon can be floated on these grids if the sample always settles on carbon and not the holes.
  • C-flat - commercially available. Thinner than Quantifoil, but more fragile. May be able to image over this carbon
  • Lacey Carbon - Home-made plastic films which are carbon coated, then plastic is dissolved away. These grids can be used to support tissue sections, or can serve as support for a very thin carbon film and used also for single particle analysis.

Uncommon coatings

• • Beryllium
  • Silicone monoxide - low background contrast, stable under beam, more hydrophilic than carbon

• Set ALLOWTOPICCHANGE = CemfacGroup

-- KdDerr - 28 Aug 2007

Grid preparation

Principles
Grid selection

Protocols
Solid Carbon Films
Triafol Holey Films
Claudio Holey Films
Carbon sandwich
Fiducials for tomography

Supplementary

Principles
Grid selection
Deciding the grids required for a particular EM application is an important but easy decision. There seems to be an overwhelming variety of possibilities. The camp can be divided in two: tissue samples for tomography or imaging of isolated proteins. Usually, proteins are imaged using 300 or 400 square mesh grids, while tissue sections can be placed on slotted grids for improved tilting capabilities.

Shiny vs. Dull side
There is no solid evidence that using either side is better, therefore as long as you keep track of which side you use, everything should be fine. The shiny side tends to be smoother, while the dull side is more like sand paper. There are grids that have one side flashed with Rh, allowing a very easy identification of the side where the sample is placed.

Mesh Size (lines per inch)
There is a great variety of mesh shapes and sizes for grids. The most common selections are as follows:

Principles

Protocols

Grid Preparation

• Solid Carbon Films
• Triafol Holey Films
• Claudio Holey Films
• Carbon sandwich
• Fiducials for tomography

Grid selection

Shiny vs. Dull side

Mesh Size (lines per inch)

• 400 mesh - recommended for support films. Protein imaging applications, for example.
• 200 mesh - A good compromise for tomography.
• Slotted grids - For tomography and serial sections. Need to be coated with a plastic film.

Material

Common

• Copper - Most common - inexpensive, conductive, beam stable. Relatively thin grid bars.
• Copper/Palladium - Palladium adds rigidity to the grid, but makes it more expensive.
• Copper/Rhodium - non-tarnishing, reduction in bar thickness. Recognizable sides.

Less common

• Gold or Platinum - non-reactive, expensive, beam stable. Easily bent (especially gold)
• Molybdenum - Expensive, Coefficient of contraction similar to carbon. Quite rigid.
• Nickel - Used mainly for silver enhancement.
• Beryllium - Minimize background radiation. Mainly used for material science.
• Titanium - Extra stiff.

Mesh types

• Square Mesh. These are by far the most common. There are many brands with different peculiarities each.
• Finder Grids - These are particularly useful when one needs to go back to collect data from a known area.
• Hexagonal Mesh - Used for tomography more than anything, but can be used for the same applications as the square grids.
• Slot Grids - For tomography. More fragile than others since there is less support

Support films

With CarbonCoating

• Formvar and Carbon - A formvar film is made on a glass slide. Then the film is floated in water and grids placed on it. The grids are then picked up with paper from the water surface and carbon is evaporated on top of the formvar.
• Carbon only - In this case, carbon is evaporated on a freshly cleaved mica sheet. The carbon film is then floated off of the mica and lowered onto submerged grids placed on a piece of filter paper.
• Nitrocellulose and Carbon - Similar to the formvar-carbon film. A drop of nitrocellulose is placed on a crystallization dish, forming thus a thin plastic film. Grids are placed on the surface, picked up with paper and coated with carbon.

• Carbon with holes - These films are used commonly for frozen hydrated samples. There are several ways to prepare them, or they can be purchased from vendors (Quantifoil, C-flat). Commercially available grids have regular holes.
  • Quantifoil - commercially available. Carbon is ~200 A thick, sturdy, but generally not suitable for imaging. A thin coating of carbon can be floated on these grids if the sample always settles on carbon and not the holes.
  • C-flat - commercially available. Thinner than Quantifoil, but more fragile. May be able to image over this carbon
  • Lacey Carbon - Home-made plastic films which are carbon coated, then plastic is dissolved away. These grids can be used to support tissue sections, or can serve as support for a very thin carbon film and used also for single particle analysis.

Uncommon coatings

• • Beryllium
  • Silicone monoxide - low background contrast, stable under beam, more hydrophilic than carbon

• Set ALLOWTOPICCHANGE = CemfacGroup

-- KdDerr - 28 Aug 2007

Grid preparation

Principles
Grid selection

Protocols
Solid Carbon Films
Triafol Holey Films
Claudio Holey Films
Carbon sandwich
Fiducials for tomography

Supplementary

Principles
Grid selection
Deciding the grids required for a particular EM application is an important but easy decision. There seems to be an overwhelming variety of possibilities. The camp can be divided in two: tissue samples for tomography or imaging of isolated proteins. Usually, proteins are imaged using 300 or 400 square mesh grids, while tissue sections can be placed on slotted grids for improved tilting capabilities.

Shiny vs. Dull side
There is no solid evidence that using either side is better, therefore as long as you keep track of which side you use, everything should be fine. The shiny side tends to be smoother, while the dull side is more like sand paper. There are grids that have one side flashed with Rh, allowing a very easy identification of the side where the sample is placed.

Mesh Size (lines per inch)
There is a great variety of mesh shapes and sizes for grids. The most common selections are as follows:

Principles

Protocols

Grid Preparation

• Solid Carbon Films
• Triafol Holey Films
• Claudio Holey Films
• Carbon sandwich
• Fiducials for tomography

Grid selection

Shiny vs. Dull side

Mesh Size (lines per inch)

• 400 mesh - recommended for support films. Protein imaging applications, for example.
• 200 mesh - A good compromise for tomography.
• Slotted grids - For tomography and serial sections. Need to be coated with a plastic film.

Material

Common

• Copper - Most common - inexpensive, conductive, beam stable. Relatively thin grid bars.
• Copper/Palladium - Palladium adds rigidity to the grid, but makes it more expensive.
• Copper/Rhodium - non-tarnishing, reduction in bar thickness. Recognizable sides.

Less common

• Gold or Platinum - non-reactive, expensive, beam stable. Easily bent (especially gold)
• Molybdenum - Expensive, Coefficient of contraction similar to carbon. Quite rigid.
• Nickel - Used mainly for silver enhancement.
• Beryllium - Minimize background radiation. Mainly used for material science.
• Titanium - Extra stiff.

Mesh types

• Square Mesh. These are by far the most common. There are many brands with different peculiarities each.
• Finder Grids - These are particularly useful when one needs to go back to collect data from a known area.
• Hexagonal Mesh - Used for tomography more than anything, but can be used for the same applications as the square grids.
• Slot Grids - For tomography. More fragile than others since there is less support

Support films

With CarbonCoating

• Formvar and Carbon - A formvar film is made on a glass slide. Then the film is floated in water and grids placed on it. The grids are then picked up with paper from the water surface and carbon is evaporated on top of the formvar.
• Carbon only - In this case, carbon is evaporated on a freshly cleaved mica sheet. The carbon film is then floated off of the mica and lowered onto submerged grids placed on a piece of filter paper.
• Nitrocellulose and Carbon - Similar to the formvar-carbon film. A drop of nitrocellulose is placed on a crystallization dish, forming thus a thin plastic film. Grids are placed on the surface, picked up with paper and coated with carbon.

• Carbon with holes - These films are used commonly for frozen hydrated samples. There are several ways to prepare them, or they can be purchased from vendors (Quantifoil, C-flat). Commercially available grids have regular holes.
  • Quantifoil - commercially available. Carbon is ~200 A thick, sturdy, but generally not suitable for imaging. A thin coating of carbon can be floated on these grids if the sample always settles on carbon and not the holes.
  • C-flat - commercially available. Thinner than Quantifoil, but more fragile. May be able to image over this carbon
  • Lacey Carbon - Home-made plastic films which are carbon coated, then plastic is dissolved away. These grids can be used to support tissue sections, or can serve as support for a very thin carbon film and used also for single particle analysis.

Uncommon coatings

• • Beryllium
  • Silicone monoxide - low background contrast, stable under beam, more hydrophilic than carbon

• Set ALLOWTOPICCHANGE = CemfacGroup

-- KdDerr - 28 Aug 2007

Grid preparation

Principles
Grid selection

Protocols
Solid Carbon Films
Triafol Holey Films
Claudio Holey Films
Carbon sandwich
Fiducials for tomography

Supplementary

Principles
Grid selection
Deciding the grids required for a particular EM application is an important but easy decision. There seems to be an overwhelming variety of possibilities. The camp can be divided in two: tissue samples for tomography or imaging of isolated proteins. Usually, proteins are imaged using 300 or 400 square mesh grids, while tissue sections can be placed on slotted grids for improved tilting capabilities.

Shiny vs. Dull side
There is no solid evidence that using either side is better, therefore as long as you keep track of which side you use, everything should be fine. The shiny side tends to be smoother, while the dull side is more like sand paper. There are grids that have one side flashed with Rh, allowing a very easy identification of the side where the sample is placed.

Mesh Size (lines per inch)
There is a great variety of mesh shapes and sizes for grids. The most common selections are as follows:

Principles

• Solid Carbon Films
• Triafol Holey Films
• Claudio Holey Films
• Carbon sandwich
• Fiducials for tomography

Grid selection

Shiny vs. Dull side

Mesh Size (lines per inch)

• 400 mesh - recommended for support films. Protein imaging applications, for example.
• 200 mesh - A good compromise for tomography.
• Slotted grids - For tomography and serial sections. Need to be coated with a plastic film.

Material

Common

• Copper - Most common - inexpensive, conductive, beam stable. Relatively thin grid bars.
• Copper/Palladium - Palladium adds rigidity to the grid, but makes it more expensive.
• Copper/Rhodium - non-tarnishing, reduction in bar thickness. Recognizable sides.

Less common

• Gold or Platinum - non-reactive, expensive, beam stable. Easily bent (especially gold)
• Molybdenum - Expensive, Coefficient of contraction similar to carbon. Quite rigid.
• Nickel - Used mainly for silver enhancement.
• Beryllium - Minimize background radiation. Mainly used for material science.
• Titanium - Extra stiff.

Mesh types

• Square Mesh. These are by far the most common. There are many brands with different peculiarities each.
• Finder Grids - These are particularly useful when one needs to go back to collect data from a known area.
• Hexagonal Mesh - Used for tomography more than anything, but can be used for the same applications as the square grids.
• Slot Grids - For tomography. More fragile than others since there is less support

Support films

With CarbonCoating

• Formvar and Carbon - A formvar film is made on a glass slide. Then the film is floated in water and grids placed on it. The grids are then picked up with paper from the water surface and carbon is evaporated on top of the formvar.
• Carbon only - In this case, carbon is evaporated on a freshly cleaved mica sheet. The carbon film is then floated off of the mica and lowered onto submerged grids placed on a piece of filter paper.
• Nitrocellulose and Carbon - Similar to the formvar-carbon film. A drop of nitrocellulose is placed on a crystallization dish, forming thus a thin plastic film. Grids are placed on the surface, picked up with paper and coated with carbon.

• Carbon with holes - These films are used commonly for frozen hydrated samples. There are several ways to prepare them, or they can be purchased from vendors (Quantifoil, C-flat). Commercially available grids have regular holes.
  • Quantifoil - commercially available. Carbon is ~200 A thick, sturdy, but generally not suitable for imaging. A thin coating of carbon can be floated on these grids if the sample always settles on carbon and not the holes.
  • C-flat - commercially available. Thinner than Quantifoil, but more fragile. May be able to image over this carbon
  • Lacey Carbon - Home-made plastic films which are carbon coated, then plastic is dissolved away. These grids can be used to support tissue sections, or can serve as support for a very thin carbon film and used also for single particle analysis.

Uncommon coatings

• • Beryllium
  • Silicone monoxide - low background contrast, stable under beam, more hydrophilic than carbon

• Set ALLOWTOPICCHANGE = CemfacGroup

-- KdDerr - 28 Aug 2007

Grid preparation

Principles
Grid selection

Protocols
Solid Carbon Films
Triafol Holey Films
Claudio Holey Films
Carbon sandwich
Fiducials for tomography

Supplementary

Principles
Grid selection
Deciding the grids required for a particular EM application is an important but easy decision. There seems to be an overwhelming variety of possibilities. The camp can be divided in two: tissue samples for tomography or imaging of isolated proteins. Usually, proteins are imaged using 300 or 400 square mesh grids, while tissue sections can be placed on slotted grids for improved tilting capabilities.

Shiny vs. Dull side
There is no solid evidence that using either side is better, therefore as long as you keep track of which side you use, everything should be fine. The shiny side tends to be smoother, while the dull side is more like sand paper. There are grids that have one side flashed with Rh, allowing a very easy identification of the side where the sample is placed.

Mesh Size (lines per inch)
There is a great variety of mesh shapes and sizes for grids. The most common selections are as follows:

Protocols

Grid selection

Shiny vs. Dull side

Mesh Size (lines per inch)

• 400 mesh - recommended for support films. Protein imaging applications, for example.
• 200 mesh - A good compromise for tomography.
• Slotted grids - For tomography and serial sections. Need to be coated with a plastic film.

Material

Common

• Copper - Most common - inexpensive, conductive, beam stable. Relatively thin grid bars.
• Copper/Palladium - Palladium adds rigidity to the grid, but makes it more expensive.
• Copper/Rhodium - non-tarnishing, reduction in bar thickness. Recognizable sides.

Less common

• Gold or Platinum - non-reactive, expensive, beam stable. Easily bent (especially gold)
• Molybdenum - Expensive, Coefficient of contraction similar to carbon. Quite rigid.
• Nickel - Used mainly for silver enhancement.
• Beryllium - Minimize background radiation. Mainly used for material science.
• Titanium - Extra stiff.

Mesh types

• Square Mesh. These are by far the most common. There are many brands with different peculiarities each.
• Finder Grids - These are particularly useful when one needs to go back to collect data from a known area.
• Hexagonal Mesh - Used for tomography more than anything, but can be used for the same applications as the square grids.
• Slot Grids - For tomography. More fragile than others since there is less support

Support films

With CarbonCoating

• Formvar and Carbon - A formvar film is made on a glass slide. Then the film is floated in water and grids placed on it. The grids are then picked up with paper from the water surface and carbon is evaporated on top of the formvar.
• Carbon only - In this case, carbon is evaporated on a freshly cleaved mica sheet. The carbon film is then floated off of the mica and lowered onto submerged grids placed on a piece of filter paper.
• Nitrocellulose and Carbon - Similar to the formvar-carbon film. A drop of nitrocellulose is placed on a crystallization dish, forming thus a thin plastic film. Grids are placed on the surface, picked up with paper and coated with carbon.

• Carbon with holes - These films are used commonly for frozen hydrated samples. There are several ways to prepare them, or they can be purchased from vendors (Quantifoil, C-flat). Commercially available grids have regular holes.
  • Quantifoil - commercially available. Carbon is ~200 A thick, sturdy, but generally not suitable for imaging. A thin coating of carbon can be floated on these grids if the sample always settles on carbon and not the holes.
  • C-flat - commercially available. Thinner than Quantifoil, but more fragile. May be able to image over this carbon
  • Lacey Carbon - Home-made plastic films which are carbon coated, then plastic is dissolved away. These grids can be used to support tissue sections, or can serve as support for a very thin carbon film and used also for single particle analysis.

• • Beryllium
  • Silicone monoxide - low background contrast, stable under beam, more hydrophilic than carbon

• Set ALLOWTOPICCHANGE = CemfacGroup

-- KdDerr - 28 Aug 2007

Grid preparation

Principles
Grid selection

Protocols
Solid Carbon Films
Triafol Holey Films
Claudio Holey Films
Carbon sandwich
Fiducials for tomography

Supplementary

Principles
Grid selection
Deciding the grids required for a particular EM application is an important but easy decision. There seems to be an overwhelming variety of possibilities. The camp can be divided in two: tissue samples for tomography or imaging of isolated proteins. Usually, proteins are imaged using 300 or 400 square mesh grids, while tissue sections can be placed on slotted grids for improved tilting capabilities.

Shiny vs. Dull side
There is no solid evidence that using either side is better, therefore as long as you keep track of which side you use, everything should be fine. The shiny side tends to be smoother, while the dull side is more like sand paper. There are grids that have one side flashed with Rh, allowing a very easy identification of the side where the sample is placed.

Mesh Size (lines per inch)
There is a great variety of mesh shapes and sizes for grids. The most common selections are as follows:

• Set ALLOWTOPICCHANGE = CemfacGroup

-- KdDerr - 28 Aug 2007

Grid preparation

Principles
Grid selection

Protocols
Solid Carbon Films
Triafol Holey Films
Claudio Holey Films
Carbon sandwich
Fiducials for tomography

Supplementary

Principles
Grid selection
Deciding the grids required for a particular EM application is an important but easy decision. There seems to be an overwhelming variety of possibilities. The camp can be divided in two: tissue samples for tomography or imaging of isolated proteins. Usually, proteins are imaged using 300 or 400 square mesh grids, while tissue sections can be placed on slotted grids for improved tilting capabilities.

Shiny vs. Dull side
There is no solid evidence that using either side is better, therefore as long as you keep track of which side you use, everything should be fine. The shiny side tends to be smoother, while the dull side is more like sand paper. There are grids that have one side flashed with Rh, allowing a very easy identification of the side where the sample is placed.

Mesh Size (lines per inch)
There is a great variety of mesh shapes and sizes for grids. The most common selections are as follows:

• Set ALLOWTOPICCHANGE = CemfacGroup

-- KdDerr - 28 Aug 2007

Grid preparation

Principles
Grid selection

Protocols
Solid Carbon Films
Triafol Holey Films
Claudio Holey Films
Carbon sandwich
Fiducials for tomography

Supplementary

Principles
Grid selection
Deciding the grids required for a particular EM application is an important but easy decision. There seems to be an overwhelming variety of possibilities. The camp can be divided in two: tissue samples for tomography or imaging of isolated proteins. Usually, proteins are imaged using 300 or 400 square mesh grids, while tissue sections can be placed on slotted grids for improved tilting capabilities.

Shiny vs. Dull side
There is no solid evidence that using either side is better, therefore as long as you keep track of which side you use, everything should be fine. The shiny side tends to be smoother, while the dull side is more like sand paper. There are grids that have one side flashed with Rh, allowing a very easy identification of the side where the sample is placed.

Mesh Size (lines per inch)
There is a great variety of mesh shapes and sizes for grids. The most common selections are as follows:

• Set ALLOWTOPICCHANGE = CemfacGroup

-- KdDerr - 28 Aug 2007

Grid preparation

Principles
Grid selection

Protocols
Solid Carbon Films
Triafol Holey Films
Claudio Holey Films
Carbon sandwich
Fiducials for tomography

Supplementary

Principles
Grid selection
Deciding the grids required for a particular EM application is an important but easy decision. There seems to be an overwhelming variety of possibilities. The camp can be divided in two: tissue samples for tomography or imaging of isolated proteins. Usually, proteins are imaged using 300 or 400 square mesh grids, while tissue sections can be placed on slotted grids for improved tilting capabilities.

Shiny vs. Dull side
There is no solid evidence that using either side is better, therefore as long as you keep track of which side you use, everything should be fine. The shiny side tends to be smoother, while the dull side is more like sand paper. There are grids that have one side flashed with Rh, allowing a very easy identification of the side where the sample is placed.

Mesh Size (lines per inch)
There is a great variety of mesh shapes and sizes for grids. The most common selections are as follows:

• Set ALLOWTOPICCHANGE = CemfacGroup

-- KdDerr - 28 Aug 2007

Grid preparation

Principles
Grid selection

Protocols
Solid Carbon Films
Triafol Holey Films
Claudio Holey Films
Carbon sandwich

Supplementary

Principles
Grid selection
Deciding the grids required for a particular EM application is an important but easy decision. There seems to be an overwhelming variety of possibilities. The camp can be divided in two: tissue samples for tomography or imaging of isolated proteins. Usually, proteins are imaged using 300 or 400 square mesh grids, while tissue sections can be placed on slotted grids for improved tilting capabilities.

Shiny vs. Dull side
There is no solid evidence that using either side is better, therefore as long as you keep track of which side you use, everything should be fine. The shiny side tends to be smoother, while the dull side is more like sand paper. There are grids that have one side flashed with Rh, allowing a very easy identification of the side where the sample is placed.

Mesh Size (lines per inch)
There is a great variety of mesh shapes and sizes for grids. The most common selections are as follows:

• Set ALLOWTOPICCHANGE = CemfacGroup

-- KdDerr - 28 Aug 2007

Grid preparation

Principles
Grid selection

Protocols
Solid Carbon Films
Triafol Holey Films
Claudio Holey Films

Supplementary

Principles
Grid selection
Deciding the grids required for a particular EM application is an important but easy decision. There seems to be an overwhelming variety of possibilities. The camp can be divided in two: tissue samples for tomography or imaging of isolated proteins. Usually, proteins are imaged using 300 or 400 square mesh grids, while tissue sections can be placed on slotted grids for improved tilting capabilities.

Shiny vs. Dull side
There is no solid evidence that using either side is better, therefore as long as you keep track of which side you use, everything should be fine. The shiny side tends to be smoother, while the dull side is more like sand paper. There are grids that have one side flashed with Rh, allowing a very easy identification of the side where the sample is placed.

Mesh Size (lines per inch)
There is a great variety of mesh shapes and sizes for grids. The most common selections are as follows:

• Set ALLOWTOPICCHANGE = CemfacGroup

-- KdDerr - 28 Aug 2007

Grid preparation

Principles
Grid selection

Protocols
Solid Carbon Films
Triafol Holey Films

Supplementary

Principles
Grid selection
Deciding the grids required for a particular EM application is an important but easy decision. There seems to be an overwhelming variety of possibilities. The camp can be divided in two: tissue samples for tomography or imaging of isolated proteins. Usually, proteins are imaged using 300 or 400 square mesh grids, while tissue sections can be placed on slotted grids for improved tilting capabilities.

Shiny vs. Dull side
There is no solid evidence that using either side is better, therefore as long as you keep track of which side you use, everything should be fine. The shiny side tends to be smoother, while the dull side is more like sand paper. There are grids that have one side flashed with Rh, allowing a very easy identification of the side where the sample is placed.

Mesh Size (lines per inch)
There is a great variety of mesh shapes and sizes for grids. The most common selections are as follows:

• Set ALLOWTOPICCHANGE = CemfacGroup

-- KdDerr - 28 Aug 2007

Grid preparation

Principles
Grid selection

Protocols
Solid Carbon Films

Supplementary

Principles
Grid selection
Deciding the grids required for a particular EM application is an important but easy decision. There seems to be an overwhelming variety of possibilities. The camp can be divided in two: tissue samples for tomography or imaging of isolated proteins. Usually, proteins are imaged using 300 or 400 square mesh grids, while tissue sections can be placed on slotted grids for improved tilting capabilities.

Shiny vs. Dull side
There is no solid evidence that using either side is better, therefore as long as you keep track of which side you use, everything should be fine. The shiny side tends to be smoother, while the dull side is more like sand paper. There are grids that have one side flashed with Rh, allowing a very easy identification of the side where the sample is placed.

Mesh Size (lines per inch)
There is a great variety of mesh shapes and sizes for grids. The most common selections are as follows:

• Set ALLOWTOPICCHANGE = CemfacGroup

-- KdDerr - 28 Aug 2007

Grid preparation

Principles
Grid selection

Protocols

Supplementary

Principles
Grid selection
Deciding the grids required for a particular EM application is an important but easy decision. There seems to be an overwhelming variety of possibilities. The camp can be divided in two: tissue samples for tomography or imaging of isolated proteins. Usually, proteins are imaged using 300 or 400 square mesh grids, while tissue sections can be placed on slotted grids for improved tilting capabilities.

Shiny vs. Dull side
There is no solid evidence that using either side is better, therefore as long as you keep track of which side you use, everything should be fine. The shiny side tends to be smoother, while the dull side is more like sand paper. There are grids that have one side flashed with Rh, allowing a very easy identification of the side where the sample is placed.

Mesh Size (lines per inch)
There is a great variety of mesh shapes and sizes for grids. The most common selections are as follows:

• Set ALLOWTOPICCHANGE = CemfacGroup

-- KdDerr - 28 Aug 2007

Grid preparation

Principles
Grid selection

Protocols
Solid Carbon Films^?
Triafol Holey Films^?
Claudio Holey Films^?
Carbon sandwich^?

Supplementary

Principles
Grid selection
Deciding the grids required for a particular EM application is an important but easy decision. There seems to be an overwhelming variety of possibilities. The camp can be divided in two: tissue samples for tomography or imaging of isolated proteins. Usually, proteins are imaged using 300 or 400 square mesh grids, while tissue sections can be placed on slotted grids for improved tilting capabilities.

Shiny vs. Dull side
There is no solid evidence that using either side is better, therefore as long as you keep track of which side you use, everything should be fine. The shiny side tends to be smoother, while the dull side is more like sand paper. There are grids that have one side flashed with Rh, allowing a very easy identification of the side where the sample is placed.

Mesh Size (lines per inch)
There is a great variety of mesh shapes and sizes for grids. The most common selections are as follows:

• Set ALLOWTOPICCHANGE = CemfacGroup

-- KdDerr - 28 Aug 2007

Grid preparation

Principles
Grid selection

Protocols
Solid Carbon Films^?
Triafol Holey Films^?
Claudio Holey Films^?

Supplementary

Principles
Grid selection
Deciding the grids required for a particular EM application is an important but easy decision. There seems to be an overwhelming variety of possibilities. The camp can be divided in two: tissue samples for tomography or imaging of isolated proteins. Usually, proteins are imaged using 300 or 400 square mesh grids, while tissue sections can be placed on slotted grids for improved tilting capabilities.

Shiny vs. Dull side
There is no solid evidence that using either side is better, therefore as long as you keep track of which side you use, everything should be fine. The shiny side tends to be smoother, while the dull side is more like sand paper. There are grids that have one side flashed with Rh, allowing a very easy identification of the side where the sample is placed.

Mesh Size (lines per inch)
There is a great variety of mesh shapes and sizes for grids. The most common selections are as follows:

• Set ALLOWTOPICCHANGE = CemfacGroup

-- KdDerr - 28 Aug 2007

Grid preparation

Principles
Grid selection

Protocols
Solid Carbon Films^?
Triafol Holey Films^?

Supplementary

Principles
Grid selection
Deciding the grids required for a particular EM application is an important but easy decision. There seems to be an overwhelming variety of possibilities. The camp can be divided in two: tissue samples for tomography or imaging of isolated proteins. Usually, proteins are imaged using 300 or 400 square mesh grids, while tissue sections can be placed on slotted grids for improved tilting capabilities.

Shiny vs. Dull side
There is no solid evidence that using either side is better, therefore as long as you keep track of which side you use, everything should be fine. The shiny side tends to be smoother, while the dull side is more like sand paper. There are grids that have one side flashed with Rh, allowing a very easy identification of the side where the sample is placed.

Mesh Size (lines per inch)
There is a great variety of mesh shapes and sizes for grids. The most common selections are as follows:

• Set ALLOWTOPICCHANGE = CemfacGroup

-- KdDerr - 28 Aug 2007

Grid preparation

Principles
Grid selection

Protocols
Solid Carbon Films^?
Triafol Holey Films^?

Supplementary

Principles
Grid selection
Deciding the grids required for a particular EM application is an important but easy decision. There seems to be an overwhelming variety of possibilities. The camp can be divided in two: tissue samples for tomography or imaging of isolated proteins. Usually, proteins are imaged using 300 or 400 square mesh grids, while tissue sections can be placed on slotted grids for improved tilting capabilities.

Shiny vs. Dull side
There is no solid evidence that using either side is better, therefore as long as you keep track of which side you use, everything should be fine. The shiny side tends to be smoother, while the dull side is more like sand paper. There are grids that have one side flashed with Rh, allowing a very easy identification of the side where the sample is placed.

Mesh Size (lines per inch)
There is a great variety of mesh shapes and sizes for grids. The most common selections are as follows:

• Set ALLOWTOPICCHANGE = CemfacGroup

-- KdDerr - 28 Aug 2007

Grid preparation

Principles
Grid selection

Protocols
Solid Carbon Films^?
Triafol Holey Films^?

Supplementary

Principles
Grid selection
Deciding the grids required for a particular EM application is an important but easy decision. There seems to be an overwhelming variety of possibilities. The camp can be divided in two: tissue samples for tomography or imaging of isolated proteins. Usually, proteins are imaged using 300 or 400 square mesh grids, while tissue sections can be placed on slotted grids for improved tilting capabilities.

Shiny vs. Dull side
There is no solid evidence that using either side is better, therefore as long as you keep track of which side you use, everything should be fine. The shiny side tends to be smoother, while the dull side is more like sand paper. There are grids that have one side flashed with Rh, allowing a very easy identification of the side where the sample is placed.

Mesh Size (lines per inch)
There is a great variety of mesh shapes and sizes for grids. The most common selections are as follows:

• Set ALLOWTOPICCHANGE = CemfacGroup

-- KdDerr - 28 Aug 2007

Grid preparation

Principles
Grid selection

Protocols
Solid Carbon Films^?

Supplementary

Principles
Grid selection
Deciding the grids required for a particular EM application is an important but easy decision. There seems to be an overwhelming variety of possibilities. The camp can be divided in two: tissue samples for tomography or imaging of isolated proteins. Usually, proteins are imaged using 300 or 400 square mesh grids, while tissue sections can be placed on slotted grids for improved tilting capabilities.

Shiny vs. Dull side
There is no solid evidence that using either side is better, therefore as long as you keep track of which side you use, everything should be fine. The shiny side tends to be smoother, while the dull side is more like sand paper. There are grids that have one side flashed with Rh, allowing a very easy identification of the side where the sample is placed.

Mesh Size (lines per inch)
There is a great variety of mesh shapes and sizes for grids. The most common selections are as follows:

• Set ALLOWTOPICCHANGE = CemfacGroup

-- KdDerr - 28 Aug 2007

Grid preparation

Principles
Grid selection

Protocols

Supplementary

Principles
Grid selection
Deciding the grids required for a particular EM application is an important but easy decision. There seems to be an overwhelming variety of possibilities. The camp can be divided in two: tissue samples for tomography or imaging of isolated proteins. Usually, proteins are imaged using 300 or 400 square mesh grids, while tissue sections can be placed on slotted grids for improved tilting capabilities.

Shiny vs. Dull side
There is no solid evidence that using either side is better, therefore as long as you keep track of which side you use, everything should be fine. The shiny side tends to be smoother, while the dull side is more like sand paper. There are grids that have one side flashed with Rh, allowing a very easy identification of the side where the sample is placed.

Mesh Size (lines per inch)
There is a great variety of mesh shapes and sizes for grids. The most common selections are as follows:

• Set ALLOWTOPICCHANGE = CemfacGroup

-- KdDerr - 28 Aug 2007

Grid preparation

Principles
Grid selection

Protocols
Solid Carbon Films^?
Triafol Holey Films^?
Claudio Holey Films^?
Carbon sandwich^?
Fiducials for tomography^?

Supplementary

Principles
Grid selection
Deciding the grids required for a particular EM application is an important but easy decision. There seems to be an overwhelming variety of possibilities. The camp can be divided in two: tissue samples for tomography or imaging of isolated proteins. Usually, proteins are imaged using 300 or 400 square mesh grids, while tissue sections can be placed on slotted grids for improved tilting capabilities.

Shiny vs. Dull side
There is no solid evidence that using either side is better, therefore as long as you keep track of which side you use, everything should be fine. The shiny side tends to be smoother, while the dull side is more like sand paper. There are grids that have one side flashed with Rh, allowing a very easy identification of the side where the sample is placed.

Mesh Size (lines per inch)
There is a great variety of mesh shapes and sizes for grids. The most common selections are as follows:

• Set ALLOWTOPICCHANGE = CemfacGroup

-- KdDerr - 28 Aug 2007

Grid preparation

Principles
Grid selection

Protocols
Solid Carbon Films^?
Triafol Holey Films^?
Claudio Holey Films^?
Carbon sandwich^?
Fiducials for tomography^?

Supplementary

Principles
Grid selection
Deciding the grids required for a particular EM application is an important but easy decision. There seems to be an overwhelming variety of possibilities. The camp can be divided in two: tissue samples for tomography or imaging of isolated proteins. Usually, proteins are imaged using 300 or 400 square mesh grids, while tissue sections can be placed on slotted grids for improved tilting capabilities.

Shiny vs. Dull side
There is no solid evidence that using either side is better, therefore as long as you keep track of which side you use, everything should be fine. The shiny side tends to be smoother, while the dull side is more like sand paper. There are grids that have one side flashed with Rh, allowing a very easy identification of the side where the sample is placed.

Mesh Size (lines per inch)
There is a great variety of mesh shapes and sizes for grids. The most common selections are as follows:

• Set ALLOWTOPICCHANGE = CemfacGroup

Grid preparation

Principles
Grid selection

Protocols
Solid Carbon Films^?
Triafol Holey Films^?
Claudio Holey Films^?
Carbon sandwich^?
Fiducials for tomography^?

Supplementary

Principles
Grid selection
Deciding the grids required for a particular EM application is an important but easy decision. There seems to be an overwhelming variety of possibilities. The camp can be divided in two: tissue samples for tomography or imaging of isolated proteins. Usually, proteins are imaged using 300 or 400 square mesh grids, while tissue sections can be placed on slotted grids for improved tilting capabilities.

Shiny vs. Dull side
There is no solid evidence that using either side is better, therefore as long as you keep track of which side you use, everything should be fine. The shiny side tends to be smoother, while the dull side is more like sand paper. There are grids that have one side flashed with Rh, allowing a very easy identification of the side where the sample is placed.

Mesh Size (lines per inch)
There is a great variety of mesh shapes and sizes for grids. The most common selections are as follows:

• Set ALLOWTOPICCHANGE = CemfacGroup

-- KakoliMitra - 12 Feb 2009