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Microscopy & Imaging JoVE (Open Access) Citable · DOI

Miniaturized Sample Preparation for Transmission Electron Microscopy

DOI: 10.3791/57310-v
What you'll learn
  • ✓ Prepare nanoliter-volume samples for transmission electron microscopy without paper blotting
  • ✓ Perform cryo-grid preparation and cell lysis macro protocols for single-cell analysis
  • ✓ Analyze membrane structures using miniaturized sample preparation and cryowriter imaging
Protocol

An instrument and methods for the preparation of nanoliter-sized sample volumes for transmission electron microscopy is presented. No paper-blotting steps are required, thus avoiding the detrimental consequences this can have for proteins, significantly reducing sample loss and enabling the analysis of single cell lysate for visual proteomics.

Difficulty
advanced
Total time
~2–4 hours per sample preparation and imaging session
Biosafety
BSL-1

Steps

1
Prepare cryo-grid for electron microscopy

Set up and condition electron microscopy grids for cryogenic sample mounting. This step establishes the substrate for nanoliter sample deposition.

▶ 00:46
2
Execute grid preparation macro routine

Run automated macro protocol to complete grid conditioning and initialization. This standardizes and accelerates grid setup before sample application.

▶ 02:42
3
Prepare single-cell lysate sample

Extract and lyse individual cells to generate nanoliter-volume lysate for proteomics analysis. Minimizes sample loss and maintains protein integrity without blotting.

▶ 03:56
4
Execute cell lysis macro protocol

Run automated lysis macro to standardize cell disruption and lysate preparation. Ensures consistent, reproducible sample processing for downstream imaging.

▶ 05:54
5
Image membrane structures by cryowriter

Acquire and visualize transmission electron microscopy images of cellular membrane structures using the miniaturized cryowriter instrument. Demonstrates protocol utility for visual proteomics.

▶ 07:32
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