mPrep/s™ capsules solve multiple specimen processing challenges for microscopy, and other applications
Less damage. Encapsulated specimens can’t be lost or damaged during processing
Efficient and consistent. Prepare multiple specimens simultaneously and identically
Less handling. Exchange reagents, embed and section for TEM. Exchange reagents and critical point dry or cryo-process for SEM
Precise specimen orientation for TEM and SEM using the mPrep/s Workstation
Accurate reagent delivery with pipettors can cut consumption by 90%
Experiment in a capsule. Use capsules as microtiter wells to perform experiments and microscopy preparation without removing specimens
Always labeled. Documentation for ISO and GLP standards is easy
An insertion tool is required to insert, remove and adjust the mPrep/s screen. There are two options:
Pipettor (such as a Gilson P200 volume) to enable reagent pipetting into mPrep/s capsules
mPrep/f couplers to prevent introducing damaging reagents into pipettors
Gilson Pipetman Neo (P200) pipettors bundled with mPrep/s capsules.
mPrep/s™ Pipettor Kit single-channel
mPrep/s™ Pipettor Kit eight-channel
mPrep/Bench to seal capsule bottoms during long reagent treatments
mPrep CPD holder when using a Tousimis critical point dryer
Reagent reservoirs to hold reagents for delivery of same reagent into 8 or 12 capsules attached to multi-channel pipettor
Microwell plates to hold reagents for delivery of different or same reagents into 8 or 12 capsules attached to multi-channel pipettor
This application note 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.
This application note 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.
This application note 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.
This application note 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.
This application note 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.
This application note illustrates the preparation of polymer specimens for cross-sectional analysis. Polymer drug delivery particles were embedded and cross-sectioned for transmission FTIR spectroscopy and EDS-SEM. The application note also shows how to extend these methods to the preparation of other soft materials including coatings, thin films and fibers.
Tissue processing and orienting specimens for sectioning is time consuming. mPrep/s capsules accurately orient specimens in seconds using the mPrep Workstation. Capsules used with pipettors enable simultaneous and controlled reagent processing.
Processing tissue for SEM can be tedious. There is potential for specimen damage due to contact with labware and critical point dryers. Orienting and mounting brittle CPD prepared tissues is challenging. With mPrep/s capsules specimens are protected and can be oriented prior to CPD processing.
Multiple cryogenic manipulations are required to rapidly freeze and orient tissue specimens for cryo-facing and SEM. With mPrep/s capsules, tissue samples are oriented prior to freezing and secured in the capsule through cryo-facing and SEM mounting.
Embedding and orienting polymers for cross sectioning is challenging: Particles are difficult to entrap and orienting fibers and films is time consuming. With mPrep/s capsules particles are easily entrapped, and fibers or films are oriented in seconds using the mPrep Workstation. Once loaded in mPrep/s capsules, multiple specimens can be simultaneously embedded.
Marques N, Strong J, Strader T, Hsia R-C (2018) Optimization of Automated Immuno EM for Both Pre- and Post-Embedding Labeling. Microsc. Microanal. 24 (Suppl 1), 2018: 1300. doi:10.1017/S1431927618006980
This proceedings paper and poster presents optimized protocols for ASP-1000 automation of immuno-labeling on grids and for en bloc tissues (pre-embedding). On-grid immuno-labeling is shown with sectioned brain, sectioned HeLa cells, and whole-mount bacteria. En bloc tissue immuno-labeling is shown with mouse brain that is also silver enhanced and then resin embedded using the ASP-1000. The authors discuss how automated labeling fits into the lab workflow by providing a “drastic reduction of hands-on time” from several days to 1-2 hours, by enabling overnight processing, and by eliminating manual specimen and grid transfers that can cause sample damage. Also discussed is that automation increased reproducibility and consistently lowered background labeling noise.
Strader TE, Stewart NR, August BK, Goodman SL (2018) Automated Rapid Preparation of Tissue Specimens for TEM Pathology. Microsc. Microanal. 24 (Suppl 1), 201:1284. doi:10.1017/S1431927618006906
This proceedings paper and presented poster shows how the ASP-1000 can accelerate the preparation of TEM pathology tissue specimens from 1-2 days to several hours with conventional chemical reagents. The poster provides automation protocols for the ASP-1000 that accelerate diffusion of processing reagents into specimens by directed fluid flow that is repeated as often as 600 times in just 5 minutes. Examples are provided for multiple tissues including kidney, skeletal muscle, cardiac muscle, liver and brain. The poster also addresses how the automated preparation and mPrep/s specimen capsules and Workstation provide an efficient workflow from specimen acquisition through microtomy that also provides traceability.
Strader TE, Stewart NR, August BK, Goodman SL (2018) A Versatile All-in-One Automated Processor for Electron Microscopy. Microsc. Microanal. 24 (Suppl 1), 2018: 1122. doi:10.1017/S1431927618006098
This proceedings paper presents how the ASP-1000 provides versatility and a workflow solution for research and clinical electron microscopy labs. The authors show the automated preparation of tissues and cell suspensions from laboratory receipt in aldehyde fixative through epoxy and acrylic resin infiltration as quickly as 45.5 minutes for kidney and 2 hours for other tissues. Additional ASP-1000 applications addressed include serial block face SEM specimen preparation, on-grid and en bloc immuno-labeling, and applying gold fiducials to grids for TEM tomography.
McClain M, Nowotarski SH, Zhao Z, Alvarado AS (2017) Rapid Automated en Bloc Staining for SEM of Sections. Microsc. Microanal. PDP-59
McClain and coauthors prepared whole planarian flatworms for serial SEM imaging by developing a rapid automated version of the serial block face SEM protocol developed at the National Center for Microscopy and Imaging Research (NCMIR), which included osmium- thiocarbohydrazide–osmium and lead aspartate staining. With the McClain method, similar results were achieved in just 1.5 days by using rapid automated reagent exchanges with the ASP, compared to 7 days of manual reagent exchanges with the NCMIR protocol. This poster presentation at Microscopy and Microanalysis 2017 also received the 2nd place Diatome Award.
Lyden T, Cole K, Goodman S (2016) Application of a New Miniature Bioreactor System to Generate and Test Artificial Tumor and Normal Breast Ductal Tissues Using MCF-7/MCF10A Cells. Experimental Biology, W282, 1300.18.
This poster shows using mPrep/s capsules as micro-bioreactors to grow 3-D artificial tumors from MCF-7 breast cancer tumor cells and normal MCF10A breast ductal tissues on biomimetic extracellular matrix substrates. Cells were grown for up to one month into 4 mm diameter by 1 cm long artificial tissues. The “tissues” were characterized with microscopy and western blots. Shed cells from the artificial tissues were collected from media flowing through mPrep/s bioreactors. Flow cytometry showed that the cancer “tumors” shed about 10x more cells with characteristics consistent of metastatic cells.
McClain M. (2016) Bringing Speed and Automation Together with the ASP1000. Microsc. Microanal. 2016. Post deadline poster #49.
McClain developed a TEM tissue fixation and embedding protocol that utilized high speed agitation on the ASP-1000 to prepare planarian worm specimens for resin curing that required only 3 hours. The results were comparable to a 7-day embedding protocol that was previously used to prepare these worm specimens.
Goodman SL, Lyden T, Li W-J, Yen T. (2016) 3D Cell Culture and Microscopy in a Capsule with Scaffolds, Tumors & Stem Cells. Microsc. Microanal. 22 (Suppl 3), 2016: 998. doi:10.1017/S1431927616005833
3D cell culture bioreactor systems were assembled with mPrep/s capsules and standard labware. MCF-7 breast cancer cells were cultured for 24 days on 3D collagenous decellularized tissue scaffolds in Prep/s capsules assembled with lab tubing, syringes and pumps. The cancer cells grew into 3D tumor-like structures. Bone marrow derived mesenchymal stem cell pellets were loaded into mPrep/s capsules and grown for 21 days. Pellets grown in mPrep/s bioreactors with continuous flow-through media exhibited a 4-fold increase in the expression of collagen II mRNA, a marker for chondrogensis, in comparison to pellets cultured using mPrep/s capsules as microwell plates.
Goodman S.L. (2015) Efficient and Documented Preparation of Pharmaceutical Particles for Correlative Microscopy Analyses using mPrep Capsule Processing Microsc. Microanal. 21 (Suppl 3), 2015: 393-4. doi: 10.1017/S1431927615002767
Using mPrep capsules, Goodman prepared two types of pharmaceutical particles for characterization. The mPrep capsules enabled simultaneous and GLP compliant sample preparation. Goodman used mPrep/g capsules to simultaneously prepare 16 grids for TEM from two formulations of siRNA drug delivery nanoparticles with four staining protocols. The grids were then analyzed for morphology and uniformity using TEM. In a separate study he simultaneously embedded four formulations of polymeric drug delivery micro-particles to create eight blocks for cross-sectioning. Thin sections were than analyzed for chemical composition using transmission FTIR and for elemental composition using EDS-SEM.
Goodman SL, Wendt KD, Kostrna MS, Radi C (2015) Capsule-Based Processing and Handling of Electron Microscopy Specimens and Grids. Microscopy Today, 23(5): 30-7. doi:10.1017/S155192951500076
Authors from three institutions prepared animal and plant tissues for TEM. They used mPrep/s capsules for fixation through embedding, and mPrep/g capsules for grid staining. Researches assessed reagent consumption and labor efficiency of mPrep processing compared to traditional processing (using vials, embedding molds and droplet staining). The findings included: mPrep/s specimen processing reduced reagent consumption ten-fold and the number of hands-on operations seven-fold. mPrep/g grid staining reduced hands-on operations six-fold.
McClain M. (2014) High Throughput Multi Parameter TEM Chemical Processing Protocol Development with the mPrep-s Capsule System: Schmidtea mediterranea. Microsc. Microanal. 20 (Suppl 3), 2014: 1288-9. doi:10.1017/S1431927614008174
McClain simultaneously prepared 16 tissue specimens for TEM using 16 protocols to establish an optimal fixation and embedding protocol. This study varied buffer molarity, en bloc stains, and embedding resins. Reagent processing was conducted simultaneously using multi-channel pipettes and microtiter plates. The author reported, “The mPrep System facilitated the controlled and consistent testing of many conditions, which greatly increased the chances of obtaining optimized specimen preservation in one processing run.”
Goodman S. and Kostrna M (2011). Reducing Reagent Consumption and Improving Efficiency of Specimen Fixation and Embedding, Grid Staining and Archiving using mPrep™ Capsule Processing. Microscopy and Microanalysis, 17 (Suppl. 2) , pp 1174-1175. doi:10.1017/S143192761100674X.
This short proceedings paper introduced the mPrep System at Microscopy and Microanalysis 2011. The paper provides an overview of the mPrep System for TEM specimen preparation and TEM grid staining. The System is compared to conventional processing for laboratory efficiency and reagent consumption.
Tetri LJ, Diffee GM, Barton GP, Braun RK, Yoder HE, Haraldsdottir K, Eldridge MW, Goss KN (2018) Sex-Specific Skeletal Muscle Fatigability and Decreased Mitochondrial Oxidative Capacity in Adult Rats Exposed to Postnatal Hyperoxia. Front. Physiol. 9. 326. doi:10.3389/fphys.2018.00326
This work used the mPrep ASP-1000 and mPrep/s capsules to prepare rat gastrocnemius muscle for TEM in order to examine mitochondrial ultrastructure in a study that examines postnatal hyperoxia as a model for premature birth effects on aerobic capacity.
Stewart NR, August BK, Strader TE, Goodman SL (2017) Fast, Walk-away, Automated Processing of Mammalian Tissue for LM and TEM. Microsc. Microanal. PDP-52
Stewart and coauthors demonstrate the automated preparation of kidney, liver, skeletal and cardiac muscle using the mPrep ASP-1000. Reagent processing of kidney was completed in just 45 minutes and liver and muscle in only 2 hours, prior to resin curing. The method provided easy non-messy specimen orientation prior to resin infiltration, and specimens that were only directly handled once - when they were oriented and inserted into labeled mPrep/s capsules.
Stewart NR, August BK, Strader TE, Goodman SL (2017) Preparing LR White Embedded Tissue with mPrep/s Specimen Capsules. Microsc. Microanal. PDP-56
Stewart et al. provides a method for the easy embedding of tissue in LR White resin for TEM immuno-labeling and related applications. The method enables the reliable location of non-osmium treated tissue after embedding, non-messy specimen orientation prior to resin infiltration, minimal specimen handling, and automated reagent processing with the ASP-1000 in less than 45 minutes prior to resin curing.
Plumley WC. (2014) Cryo-Planing of Small Biological Specimens for SEM Using mPrep Capsules. Microsc. Microanal. 20 (Suppl 3), 2014: 1442-3. doi:10.1017/S1431927614008940
Plumley developed a fast and easy cryo-planing method to prepare animal and plant tissues for SEM using the mPrep System. Cryo-planing (also call cryo-facing) provides a unique 3D view of internal and surface features by cutting a cross-section through frozen tissue. Cryo-planing has not been widely used due to the difficulty of handling specimens throughout the tissue preparation process. Plumley used the pinch holding function of mPrep/s screens with modified mPrep/s capsules to hold and orient tissue during this process of rapid freezing, cryo-microtomy, freeze-drying and SEM mounting. He reported that this procedure “enabled easier, faster preparation of multiple small samples, while maintaining the excellent morphology and sectioned view… that cryo-planing is best known for…”
Strader TE, Goodman SL. (2015) Automated, Programmable Processing of Specimens and Grids with the mPrep ASP-1000. Microsc. Microanal. 21 (Suppl 3), 2015: 11-2. doi: 10.1017/S1431927615000859
Strader and Goodman introduce the mPrep Automated Specimen Processor (ASP), used for simultaneous sample preparation using mPrep capsules. The ASP-1000 prepares TEM and SEM specimens, and stains and labels TEM grids. The paper discusses the ASP-1000 features, including flexibility to efficiently perform routine and complicated tasks. The paper includes pictures of the instrument and examples of heart and skin tissue specimens prepared using mPrep capsule processing.
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
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 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
This tutorial demonstrates how to transfer mPrep capsules to a critical point dryer. The transfer is shown with an mPrep CPD holder designed for the Tousimis Samdri. Both mPrep/s and mPrep/g capsules are shown.
4 minutes 7 seconds
This webinar introduces and demonstrates the mPrep System. Webinar covers how the mPrep System works. It discusses the benefits of the mPrep System over current practice. And it addresses multiple TEM and SEM applications: tissues, macromolecules, immuno, nanoparticles, cryo-planing.
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.
How to use the mPrep CPD holder for specimen and grid sample preparation using the critical point method.
Instructions for using the mPrep capsule grid box for storage and labeling of mPrep/g and mPrep/s capsules.