Problems

Tissue prep for TEM can require dozens of reagent exchanges per specimen, using laboratory equipment not well suited for this purpose.  This presents microscopists with multiple challenges:

• Difficulty achieving uniform processing times across multiple samples
• Reagent consumption excessive relative to the amount required to ensure reaction
• Achieving proper orientation is time consuming
• Specimens damage from contact with air, vials, and transfer pipettes
• Lost specimens when difficult to see or they stick to containers

mPrep Solutions

• Automated walk-away reagent processing with the mPrep ASP-1000
• Simultaneous processing of reagent exchanges assures uniform timing
• Controlled reagent usage with pipettors = reduced reagent consumption
• Accurate orientation in seconds using the mPrep Workstation
• Encapsulated specimen reduces handling = fewer damaged specimens
• Entrapped specimens are harder to lose. Will not float.
• Labeled capsules are easy to track.

Problems

Staining tissue sections on grids requires using fine forceps to transfer grids into grid boxes, onto stain droplets or staining apparatuses, back into grid boxes, and eventually onto the TEM stage. As a result, microscopists experience multiple challenges:

• Grids are easily damaged
• Handling grids is tedious
• Grids are easily lost or mixed up
• Difficulty achieving uniform staining times across multiple grids
• Stains exposed to air can be contaminated, precipitate, and evaporate

mPrep solutions

• Encapsulated grids reduce handling and damage.
• Grids may only be directly handled at the microtome and the microscope
• Grids in mPrep/g capsules are always labeled
• Simultaneous processing assures uniform timing
• Enclosed capsules reduce and can even eliminate air exposure

Learn more about staining tissue sections on TEM grids in the application notes below.

Problems

Negative stain specimen preparation requires fine forceps to transfer filmed grids across droplets of macromolecule or particle suspensions, negative stains, and rinses.  This presents microscopists with multiple challenges:

• Grids and support films are easily damaged
• Handling grids is tedious
• Grids are easily lost or mixed up
• Limited quantities of specimen solutions may be available
• Specimen and reagent solutions may evaporate, precipitate or be contaminated
• Inconsistent adsorption and staining times across multiple grids

 mPrep  solutions

• Encapsulated grids reduce handling and  damage
• Grids with specimens may require direct handling only at the microscope
• Grids in mPrep/g capsules are always labeled
• Capsules require only 35 μl to prepare two grids
• Enclosed capsules reduce or eliminate air exposure
• Simultaneous processing assures uniform timing

Problems

Virus sample preparation for TEM requires using fine forceps to transfer filmed grids throughout the process.  Grids boxes may require decontamination prior to transfer to the TEM.

• Grids and support films are easily damaged
• Grids are easily lost or mixed up
• Sharp forceps are a puncture risk to protective clothing
• Grid handling is especially difficult wearing BSL3/4 positive pressure suits
• Limited quantities of specimen solutions may be available
• Specimen and reagent solutions exposed to air may evaporate, precipitate or be contaminated
• Grids and grid storage containers require decontamination
• Adsorption and staining times may be inconsistent across multiple grids

mPrep solutions

• Encapsulated grids reduce handling and damage
• Grids in mPrep/g capsules are always labeled
• Forceps are not used for virus preparation
• Direct handling of grids may require handling only at the microscope
• Capsules require only 35 μl to prepare two grids
• Enclosed capsules reduce or eliminate air exposure
• Grids in capsules are decontaminated as a unit
• Simultaneous processing assures uniform timing

Problems

Immunolabeling tissue sections on TEM grids using “droplet” methods requires the timed transfer of multiple grids through a dozen reagents in a process that can take hours or even a full day. As a result, microscopists experience multiple challenges:

• Difficultly achieving consistent reagent timing, which limits experimental reproducibility
• Grids are easily damaged, lost, or mixed up
• IGL reagents exposed to air can evaporate, precipitate or become contaminated
• IGL reagents and antibodies are expensive

mPrep solutions

• The mPrep System addresses these challenges:
  • Simultaneous processing assures identical reagent timing and reproducible results
  • Encapsulated grids are identified and protected from loss or damage
  • Enclosed capsules limit air exposure
  • mPrep/g capsules require only 20 ul per grid

Automated IGL with the mPrep ASP-1000

With the ASP-1000 the entire immunolabeling process is automated. Simply set up the reagents, start the program, and walk away for the rest of the day or overnight. Use with mPrep/g capsules for grids, and mPrep/s capsules for tissue en bloc IGL.

Manual IGL with a multi-channel pipettor

Using a multichannel pipettor ensures that reagent incubation timing is identical for all grids. Because grids are safely encapsulated reagent exchanges are fast and easy. Use with mPrep/s capsules for tissue en bloc IGL.

Problems

For successful image reconstruction colloidal gold fiducial markers must be distributed uniformly over both sides of the grid. The common protocol is to apply a droplet of a colloidal gold suspension to each grid, and then wick off the droplet after a short adsorption time. The result is that microscopists experience multiple challenges:

• Inconsistent results from grid to grid
• Uneven particle distributions on each grid and each grid side
• Time consuming to prepare multiple grids
• Grids are easily damaged, lost, or mixed up

mPrep solutions

The mPrep System addresses these challenges:

• Consistent results on every grid
• Uniform particle distribution on both grid sides
• Automated or manual simultaneous preparation of up to 16 grids
• Encapsulated grids are protected from damage or loss, and are always identified

Automated with the mPrep ASP-1000

Application of colloidal gold fiducial particles onto grids is fully automated with the mPrep ASP-1000. Colloidal gold particles are applied simultaneously to both sides of up to 16 grids. Automated agitation ensures consistent results.

Manual IGL with a multi-channel pipettor

Using mPrep/g capsules with an 8-channel pipettor, colloidal gold fiducial particles are applied simultaneously to both sides of up to 16 grids. With a multichannel pipettor, manual agitation ensures consistent results for multiple grids prepared simultaneously.

Problems

Nanoparticle specimen prep requires  forceps to transfer filmed grids through multiple reagents. Grids are stored in grid boxes until TEM imaging.

• Grids and support films are easily damaged
• Grids are easily lost or mixed up
• Limited quantities of specimen solutions may be available
• Specimen and reagent solutions may evaporate, precipitate or be contaminated
• Adsorption and staining times may be inconsistent across multiple grids

mPrep solutions

• Encapsulated grids reduce handling and damage
• Grids in mPrep/g capsules are always labeled
• Capsules require only 35 μl to prepare two grids
• Enclosed capsules reduce or eliminate air exposure
• Simultaneous processing assures uniform timing