Ultrastructural labs

A highly detailed illustration of a lab environment featuring Transmission Electron Microscopy equipment, along with sample preparation tools and ultrathin sections of biological specimens.

Ultrastructural Lab Techniques Quiz

Test your knowledge of Transmission Electron Microscopy (TEM) and its intricate sample preparation processes in this comprehensive quiz. Whether you're a student or a professional, challenge yourself with 50 questions designed to deepen your understanding of lab techniques.

Explore various stages of sample preparation.
Learn about fixation, dehydration, infiltration, and embedding.
Assess your knowledge on staining and sectioning techniques.

50 Questions12 MinutesCreated by PreparingScientist42

TEM

A technique to study structure of organisms, tissue, cells and organelles

Can study individual proteins

Forms 3D images

Sample must be coated

Electrons reflect from surface

How many steps are there in TEM sample preparation

Steps of sample preparation in TEM. (________ ______ ________ ________ ________ _______)

Size of tissue obtained for TEM

1mm

30-60nm

0.5mm

1µm

Size of section obtained for TEM

1mm

30-60nm

0.5mm

1µm

The goal of fixation is

Stopping metabolism and preventing autolysis

Replacing water content with organic solvents

Replace dehydration sol. By intermediate sol.

Make sample hard enough for cutting

Crosslinkage

Getting ultrathin sections for electron beam to cross

Transferring the sample to the tip of labeled beam capsule that was filled with resins

Dehydration is

Stopping metabolism and preventing autolysis

Replacing water content with organic solvents

Replace dehydration sol. By intermediate sol.

Make sample hard enough for cutting

Crosslinkage

Getting ultrathin sections for electron beam to cross

Transferring the sample to the tip of labeled beam capsule that was filled with resins

Infiltration is

Stopping metabolism and preventing autolysis

Replacing water content with organic solvents

Replace dehydration sol. By intermediate sol.

Make sample hard enough for cutting

Crosslinkage

Getting ultrathin sections for electron beam to cross

Transferring the sample to the tip of labeled beam capsule that was filled with resins

Embedding is

Stopping metabolism and preventing autolysis

Replacing water content with organic solvents

Replace dehydration sol. By intermediate sol.

Make sample hard enough for cutting

Crosslinkage

Getting ultrathin sections for electron beam to cross

Transferring the sample to the tip of labeled beam capsule that was filled with resins

Cross linking proteins molecules with nearby mol.

Cryofixation

Chemical fixation

Transforms water content to ice to keep structures inbplace

Done by gluteraldheyde and osmium tetraoxide

Rapid freezing of sample

Cryfixation

Chemical fixation

Done using liquid helium and nitrogen

Done using gluteraldehyde

In cryofixation the internal structure is preserved by

Cross linkage of protein mol.

Liquid helium or nitrogen

Ice

Steps in fixation

Gluteraldehyde 2.5%

Phosphate buffer

Osmium tetraoxide 1%

Water

Osmium tetraoxide

Fixes protein but damages cell membrane

Fixes cell membrane but damages protein

Gluteraldehyde

Fixes protein but damages cell membrane

Fixes cell membrane but damages protein

Appears as precipitate of fixative

Glucose inclusions

Vacuoles

Artefact

Done for 2h at 7.2 pH and 4C

Gluteraldehyde

OsO4

Embedding

Done for 45-60min at 7.2 pH and at room temp.

Gluteraldehyde

OsO4

Embedding

Done for 1-2days at 60-70C then cooled 24h

Gluteraldehyde

OsO4

Embedding

Steps of staining

Uranyl acetate

For 5 to 10 min

Water 3 changes

Lead citrate

For less than 10 min

Water

Dry on filter papre

Cutting the sample into very thin sections is called .............. . (term other than sectioning)

Break glass until you obtain triangular pieces with a sharp and a bout edge

Move and separate sections with

Forceps

Finger

Eyelash

Semi thin section

Stained with toduline blue

Studied under LM

750-850nm

All of the above

This process is called

This process is the preparation of

Specimen stage in TEM

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Halfway down

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