Advanced Microscopy and DNA Analysis Techniques: TEM, AFM, FRET, and Gel Electrophoresis

Transmission Electron Microscopy (TEM)

Technology uses a beam of elections and a detector to create images using electron scattering

Magnetic Resonance Force Microscopy

Uses a probe with a magnetic tip to manipulate the spins of the subatomic particles. The resulting vibrations are used to generate images

Fluorescense Resonance Energy Transfer

Detects energy between two fluorescent molecules, or fluorophores, when one fluorophore transfers energy to the other.

Photo Activated Localization Microscopy

Designed to detect light emitted by fluorescent proteins returning to a ground state

What were the achievements of TEM

Discovery of Viruses

Viral Structure and Life Cycle

characterizing antiparticles

Nanomedicine

Does Tem use light?

no

Which process uses a silycon probe and a lazer?

AFM

What type of images can AFM create

3D images

What led to the invention of AFM

Invention of the scanning tunneling microscope

What is the main function of palm?

absorbing and emitting

Accomplishments of PALM

Cellular Architecture and processes along with Molecular Organization

What is the main function of FRET?

Detecting energy transfers

What are the images produced by FRET?

not "real" images, but simulated images based on the calculated locations

Accomplishments of FRET

allowed scientists to measure protein-protein interactions inside of cells

X-Ray Crystallography

process of using x-rays to look at DNA samples

Accomplishments of X-Ray Crystallography

The first high-res photo of a DNA fiber (Exposure 51, with a double helix structure)

MRFM process

Detects small forces through magnetic movements (Based on AFM)

What is the Gel electrophoresi technique used for?

sorting the DNA strands according to length and comparing DNA samples using restriction enzymes.

What is the purpose of the power supply?

Provides the electrice current in order to push DNA through the gel filter

The TBE buffer solution is used to ...

help the electrical charges flow through the gel without drying up

Shorter strands of DNA travel ____ on the agarose gel than do larger strands.

farther

Which wire is closest to the well?

Black/negative wire

Which wire is farthest from the well?

red/positive wire

Do you think you would find the largest or the smallest fragment of DNA closest to the well? Explain.

You would find the largest DNA fragment closer to the well since it does not move as quickly as the smaller fragments due to its size.

What is Gel Electrophoresis

Technique for comparing DNA samples using restriction enzymes

Describe the process

DNA is cut by electric field through the gel matrix then an agarose gel is prepared, mixing the DNA with the loading buffer, loading them into the gel. Once the electric field is applied the smaller fragments to move faster and farther through the gel. The separated molecules are then visualized by using UV light to determine their sizes.