Combining Laser Capture Microdissection and Proteomics Techniques

Abstract

Laser microdissection is an effective technique to harvest pure cell populations from complex tissue sections. In addition to using the microdissected cells in several DNA and RNA studies, it has been shown that the small number of cells obtained by this technique can also be used for proteomics analysis. Combining laser capture microdissection and different types of mass spectrometers opened ways to find and identify proteins that are specific for various cell types, tissues, and their morbid alterations. Although the combination of microdissection followed by the currently available techniques of proteomics has not yet reached the stage of genome wide representation of all proteins present in a tissue, it is a feasible way to find significant differentially expressed proteins in target tissues. Recent developments in mass spectrometric detection followed by proper statistics and bioinformatics enable to analyze the proteome of not more than 100-200 cells. Obviously, validation of result is essential. The present review describes and discusses the various methods developed to target cell populations of interest by laser microdissection, followed by analysis of their proteome.

Breast tumor metastasis: analysis via proteomic profiling

“…In this way, epithelia can be separated from stromal tissue, and malignant cells can be separated from benign cells based on cytomorphology or by antibody flagging protocols [42]. The successful extraction of DNA, RNA and proteins from the microdissected material has been achieved [43,44]. Limitations of the method include the yield obtainable from minimal amounts of material and the fragmentation of the target molecules through fixation and tissue processing.…”

The challenge of gene expression profiling in heterogeneous clinical samples

“…This way of target cell isolation permits rapid, reliable laser microdissection to collect specific cell populations from a section of complex, heterogeneous tissue [89]. There are several membranes and caps used with laser capture microdissection (LCM) [90]. Dependent on the microlaser dissection device used, the collection caps used are positioned in different ways.…”

Proteomics, a new tool to monitor cancer therapy?

“…Ideally the tumour cells of the patient are obtained by laser capture microdissection. Th is method allows separation of malignant and normal cell populations within a single biopsy specimen [21]. After this approach, a variety of patient samples can be immobilized on a single array and then incubated with one detection protein or antibody to measure the expression of a single target across the arrayed cohort.…”