10 Jul Biological tissue fixation, embedding and orientation
The mPrep System streamlines the entire tissue preparation and grid staining workflow.
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.
Cardiac muscle (TEM)
Skin (TEM)
Application Notes
Demonstrates the reproducibility of rapid processing with the ASP-1000. Twenty-four (24) specimens from the same aldehyde perfusion fixed liver were prepared for TEM. Specimens were directly handled only once, when they were orientated in mPrep/s capsules. Automated processing required only 133 minutes from post-fix rinse through osmium fixation and 100% resin infiltration. All 24 samples showed well-preserved ultrastructure homogeneity as verified by TEM imaging.
Illustrates mammalian kidney tissue prep for TEM from fixation through embedding. This general protocol applies to other specimens that require embedding for cross-sectioning. Photos show the mPrep capsule reagent exchange process and how mPrep TEM blocks are faced and sectioned while still in capsules. Supplies and the reagent formulae are listed.
Demonstrates using agar embedding of suspended cells to enable rapid automated preparation with the ASP-1000 Automated processor to fix, post-fix and embed cells in epoxy for TEM. The example uses yeast, but this method can be readily extended to the preparation of other types of cells in suspension, and for the preparation of other small specimens. By combining agar embedment with ASP-1000 processing, 18 centrifugation and resuspension cycles were eliminated from a typical manual protocol. Once the cells were agar embedded, the total processing time prior to epoxy resin curing was just 1.5 hours.
Presents a method for processing mammalian muscle tissue for TEM in just over two hours. Using the ASP-1000 automated specimen processor, specimens were processed from postfixation rinse through infiltration in just 133 minutes. Specimens were handled only once, when they were oriented and inserted into mPrep/s capsules using the mPrep/s Workstation.
Presents a fast, fully-automated method of tissue processing. Mammalian kidney was prepared for TEM with the mPrep ASP-1000 programmed for highly efficient rapid-agitation. Reagent processing from postfix rinse through 100% resin infiltration was accomplished in just 45 minutes.
Required
mPrep/s capsules to hold specimens
An insertion tool is required to insert, remove and adjust the mPrep/s screen. There are two options:
- mPrep/s Insertion tool
- mPrep/s Workstation (w/integrated insertion tool): required to orient specimens using the pinch method. Recommended because of its additional benefits
- Precise specimen orientation in seconds
- Wet samples easy to handle
- Single focal plane keeps specimens in focus
- Simultaneous loading of capsules onto pipettor for reagent processing
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
mPrep/bench to seal capsule bottoms during long reagent treatments and resin curing
Recommended
Reagent reservoirs
Microwell plates
Gilson Pipetman Neo (P200) pipettors bundled with mPrep/s capsules
mPrep/s™ Pipettor Kit single-channel
- Single-channel pipettor
- mPrep/s capsule pack
- 96 pipette tips
mPrep/s™ Pipettor Kit eight-channel
- Eight-channel pipettor
- mPrep/s capsule pack
- 96 pipette tips
Publications
Rapid Automated en Bloc Staining for SEM of Sections (Microscopy & Microanalysis 2017)
Reducing Reagent Consumption and Improving Efficiency of Specimen Fixation and Embedding, Grid Staining and Archiving using mPrep™ Capsule Processing (Microscopy & Microanalysis 2011)
High Throughput Multi Parameter TEM Chemical Processing Protocol Development with the mPrep-s Capsule System: Schmidtea mediterranea (Microscopy & Microanalysis 2014)
Sex-Specific Skeletal Muscle Fatigability and Decreased Mitochondrial Oxidative Capacity in Adult Rats Exposed to Postnatal Hyperoxia (Frontiers Physiology 2018)
Bringing Speed and Automation Together with the ASP-1000 (Microscopy & Microanalysis 2016)
Capsule-Based Processing and Handling of Electron Microscopy Specimens and Grids (Microscopy Today 2015)
Automated Preparation of Core Needle Biopsy Specimens for TEM Imaging (Microscopy & Microanalysis 2019)
Preparing LR White Embedded Tissue with mPrep/s Specimen Capsules (Microscopy & Microanalysis 2017)
Fast, Walk-away, Automated Processing of Mammalian Tissue for LM and TEM (Microscopy & Microanalysis 2017)
Optimization of Automated Immuno EM for Both Pre- and Post-Embedding Labeling (Microscopy & Microanalysis 2018)
Automated Rapid Preparation of Tissue Specimens for TEM Pathology (Microscopy & Microanalysis 2018)
A Versatile All-in-One Automated Processor for Electron Microscopy. Microscopy & Microanalysis 2018)
Videos
This video tutorial shows how to trim and face an embedded block inside an mPrep/s capsule. The capsule is clamped into a microtome chuck for trimming, facing and sectioning.
1 minutes 17 seconds
The tutorial starts by introducing the mPrep Workstation. Then it demonstrates how to load and orient specimens into mPrep capsules. Three methods are shown: pinch, compression and cup.
5 minutes 19 seconds
This 18-minute webinar (broadcast 25 September 2019) presents how the ASP-1000 automates the preparation of biological specimens for TEM and SBEM (serial block face SEM) to provide walk-away convenience, reduced processing time, and easy clean-up. The webinar also discusses how the ASP-1000 provides pre-programmed protocols for preparing multiple tissues and for immunolabeling, and how users can easily modify existing or create new protocols.
Instructions
A single page graphical display of the three steps to insert specimens into mPrep/s capsules using the mPrep/s insertion tool. Use this guide as a lab reference poster.
A single page graphical display of the three steps to orient and load specimens into mPrep/s capsules. Both compression and pinch methods are shown. Use this guide as a lab reference poster.
Instructions for using mPrep/s capsules for specimen handling and processing.
Print this 96-well plate template to organize protocols into columns and rows with a microtiter 96-well plate.
This technical note introduces using mPrep capsule labeling for tracking specimens and grids with a data management system. Topics addressed include mPrep labels, 2D datamatrix barcodes, barcode readers and laboratory information management systems.
mPrep specimen products
mPrep/s™ Insertion Tool
Inserts, removes and adjusts screens in mPrep/s capsules
Product SKU: 32010$25.00
mPrep/s™ Workstation
Prepare specimens and insert into mPrep/s capsules with or without orientation using integrated screen insertion tool
Product SKU: 42100$950.00
mPrep™ CPD Holder
Holds up to 6 mPrep/s or mPrep/g capsules in Tousimis® CPD apparatus
Product SKU: 33010$225.00
ASP-1000 mPrep Automatic Specimen Processor
Programable specimen and grid preparation. Request a Quotation
Product SKU: ASP1000
mPrep/s™ Pipettor Kit eight-channel
eight-channel pipettor, mPrep/s capsule pack, 96 pipette tips
Product SKU: 43250$686.00
mPrep/s™ Pipettor Kit single-channel
single-channel pipettor, mPrep/s capsule pack, 96 pipette tips
Product SKU: 43150$366.00
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