07 Jul Negative staining of macromolecules

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The mPrep system simplifies the preparation of macromolecular and particulate specimens for TEM.

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

DNA RecA (TEM negative stain)

Application Notes

AN503: Preparing Macromolecular Specimens for TEM: Negative Staining of DNA-RecA Complex

Illustrates the preparation of macromolecular specimens for negative stain TEM using mPrep/g capsules, pipettors and Formvar®-filmed TEM grids. The example is a DNA-RecA complex, although the protocol is applicable to many other specimens. The specimen preparation and microscopy was performed in the Department of Biochemistry at the University of Wisconsin – Madison by Mr. Jay Campbell and Dr. Sindhu Chitteni-Pattu, in the laboratory of Dr. Michael M Cox.

Required

mPrep/g capsules to hold TEM grids

mPrep/f couplers to prevent introducing damaging reagents into pipettors

Pipettor (such as a Gilson P200 volume) to enable reagent pipetting into mPrep/s capsules

Product Descriptions

mPrep/g Capsules product description

Publications

Easier and Safer Biological Staining: High Contrast UranyLess Staining of TEM Grids using mPrep/g Capsules (Microscopy & Microanalysis 2015)

Abstract

Videos

Specimen and Grid Reagent Processing for TEM and SEM

This video tutorial demonstrates how to use mPrep capsules with multi-channel pipettors and common lab ware to simultaneously deliver reagents to multiple specimens and grids. The video demonstrates TEM fixation and embedding. Reagent processing procedures are similar for all specimen types and for grids with mPrep capsules .

6 minutes 4 seconds

Instructions

mPrep/g Instructions

Instructions for using mPrep/g capsules for grid handling and processing.

Template for laying out reagents

Print this 96-well plate template to organize protocols into columns and rows with a microtiter 96-well plate.

Quick Guide mPrep/g

A graphical display illustrating how to load grids into mPrep/g capsules, and how to use pipettors to stain or label grids. Use this guide as a lab reference poster.