Protein of Pure Human Breast Cancer Cells Isolated by LCM from FFPE Sample

Description

Sample Basic Information

Sample ID	Organ	Pathology Diagnosis	Gender	Age	Grade	TMN	IHC Data	Sample Format
02001T	Human breast	Human breast invasive ductal carcinoma	Female	45	2	N/A	ER(80% +), PR(70%+), E-cad(+), P120(+), CK5/6(-), CK8/18(+), Ki67(15% +), Her2 (-)	FFPE

Tissue H&E Staining and IHC images

H&E Stain

Ki 67 IHC

Flowchart of LCM  for The Product Preparation

1. FFPE samples are cut at 6 microns and mounted onto pen membrane glass slides and let the slides dry for 2 hours at room temperature.
2. After deparaffinization and rehydration, quick H&E staining is performed.
3. Arcturus XT Laser Microdissection System Tissue is used for LCM. Microdissection is performed using IR and UV laser and CapSure Macro LCM Cap.
4. The microdissected cells are on the film of Macro LCM Cap. The total dissected cells may be varied between 10,000 to 50,000 cells from different tissue type. 
5. Extract protein from LCM Cap.

Quality Control

• The tissue H&E staining slides were examined by certified pathologists. Pathological re-confirmation report is generated and digital image captured. 
• LCM is supervised by our certified pathologist.
• Three example LCM images including Before, After LCM, and LCM cap images from LCM tissue slides are took to demonstrate the dissected cells.

Extraction Protocol for Protein from LCM Caps

• For protein extraction, you can follow the protocol from Longuespée R, et al. A laser microdissection-based workflow for FFPE tissue microproteomics: Important considerations for small sample processing, Methods;2016, 104: 154-162

Product Format and Size:  

         Protein in 50 μl of lysis buffer from 2 LCM caps.

Applications

• Protein  from pure cell population microdessected from FFPE tissues can be used for proteomics.
• The product is only used for biomedical research. It is not used for clinical diagnosis.

Storage

        Store at -20ºC for shot time and  at -80ºC for longer time

References

1. Longuespée R, et al. A laser microdissection-based workflow for FFPE tissue microproteomics: Important considerations for small sample processing, Methods;2016, 104: 154-162
2. Cai J, et al. The Use of Laser Microdissection in the Identification of Suitable Reference Genes for Normalization of Quantitative Real-Time PCR in Human FFPE Epithelial Ovarian Tissue Samples. PLoS ONE 2014, 9(4): e95974. 
3. Drummond ES, et al. Proteomic analysis of neurons microdissected from formalin-fixed, paraffin-embedded Alzheimer’s disease brain tissue.Sci Rep. 2015;5:15456
4. Bockmeyer CL, et al. Recommendations for mRNA analysis of micro-dissected glomerular tufts from paraffin-embedded human kidney biopsy samples. BMC Mol Biol. 2018;19(1):2
5. Murray, Graeme I. (Ed.). Laser Capture Microdissection (Methods and Protocols, © 2018