LCM experiments have applications for (1) pathologic analysis of sections (2) analysis of lesion extent (3) clinical sample collection and reference.

Operation method

LCM Experiment

Principle

This technology uses laser cutting and other techniques to process samples that are not easily observed and detected, such as slices of biological samples, and is characterized by simplicity, speed, and high efficiency.

Materials and Instruments

RNaseAway
RNaseAway
Tape LCM Coffin Dehumidifier Dispenser Tweezers LCM System Pix Cell II Tubes without RNAase

Move

I. Materials

1. Buffers, solutions and reagents

RNaseAway (7000, MolecularBioProducts, SanDiego, CA)

2. special equipment .

Tape .

LCM coffin (Arcturus Engineering)

Dehumidifier

Dispenser, P200, P2

Tweezers

LCM System

Pix Cell II (ArcturusEngineering)

RNAase-free tubes, 0.5 ml

II Methods

According to the manufacturer's operating instructions, a brief description is given below.

1. Place the sample slice on the rotary table of the Pix Cell II device.

2. Manually position the slice to locate the transfer area.

3. Open the vacuum chuck to hold the slice in place.

4. Place the LCM cap onto the sheet by rotating the placement arm.

5. Rotate the laser interlock switch and wait 10s.

6. Press the laser button.

7. Adjust the focus by high magnification first.

The laser beam should be positioned in the center of the field of view. The laser beam through the microscope is not visible to the naked eye, so micro-separation should be done through the display when the laser beam starts working. Select the smallest point (7.5um) outside the tissue area for the test pulse to correct the focus of the laser beam.

8. Use the joystick to remove other tissues and cells beyond the target.

Immediately after LCM, the LCM cap should be removed and inserted into a 0.5 ml tube containing RNA extraction solution without RNAase.

Caveat

The LCM workspace should be clean, dry, and free of RNAase. Dispensers, forceps, and other equipment should be washed in RNAase removal solution. If humidity is high, a dehumidifier is also necessary. To obtain a good sample, the PixCelin laser beam must be properly focused. Because differences in the thickness of tissue samples can affect the focus of the laser, the tissue samples should be of uniform thickness.

Common Problems

1. Two replicates each of 0.1 cm2 and 0.3 cm2 of tissue were cut using LCM for each section. (Non-stained cuts: adjacent non-stained sections were cut using the stained section as a control).

2. Cut tissue slices were placed in a 1.5mL centrifuge tube and stored at -80℃.

This experiment is from "PCR Laboratory Guide (Second Edition)" by Seed Kang and Qu Lijia.

For more product details, please visit Aladdin Scientific website.

https://www.aladdinsci.com/