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Alzheimer Protein Research

Collect pure starting material for protein analysis. Improve your Alzheimer Protein Analysis with Laser Microdissection.

Many brain diseases result from protein malfunction, misfolding and agglutination. For these reasons protein analysis is the key to understanding the causes of, and discovering therapies for, many cerebral defects such as Alzheimer's disease.

Leica LMD systems help you easily obtain pure sufficient sample quantities from your specimen under visual control. Simply identify your regions of interest and directly cut the areas with a guided laser beam for downstream analysis. Areas can be distinct brain regions, single cells or even sub-cellular structures. You can also dissect Alzheimer’s plaques, as depicted in the image, and analyze them by mass spectrometry.

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Laser Microdissection enables users to isolate specific single cells or entire areas of tissue.

Alzheimer protein analysis workflow with automatic plaque identification and excision

Leica's Laser Microdissection systems will improve your workflow by automatically identifying and dissecting your ROIs.

This delivers sufficient and pure starting material for proteomics easily. Everything under visual control and without contamination by surrounding tissue.

1. Sample preparation

Typically, special membrane-based slides are used for laser microdissection. Sample preparation for LMD is straightforward, and can be derived from classical preparations for histology.

For downstream proteomics we recommend PET membrane slides (which are almost free of softeners) or DIRECTOR™  slides which allow completely membrane-free ablation.

Paraffin embedding brings the tissue sample into a constitution which can be cut into thin slices. Alternatively you can utilize cryo-sectioning to prepare your slides.

We will deliver the suitable LMD slides for your application.

2. Fixation and staining

Amyloid plaques can be visualized by DAB staining based on a β-amyloid antibody. This results in a brown staining of amyloid plaques in brightfield microscopy (right). DAB can be also combined with cresyl violet staining.

An alternative is Congo red which stains amyloid on formalin-fixed, paraffin-embedded (FFPE) as well as frozen tissue sections.

β-amyloid antibodies can also be used for immunofluorescence microscopy which is also applicable with Leica LMD systems.

3. Visualization and ROI

Leica LMD systems can generate an automatic sample overview that can be used to easily navigate to your regions of interest (ROI).

Your ROIs can be identified and marked automatically by the AVC software module, or you can apply shapes manually. 

4. Laser microdissection

Next, the defined ROIs are precisely dissected by the laser under visual control and directly collected via gravity.

Isolate disease affected material or any other ROI with a few mouse clicks and pool as much as you want in one or different reaction vessels if desired.

Also, you can cut your samples directly on the fly with the "Move and Cut" tool, without any predefined shapes.

The collection via gravity allows the use of standard, cost-effective consumables such as PCR tubes or 8-strip tubes.

5. Extraction of Proteins

Proteins can be extracted and prepared for downstream analysis from the dissectates with established protocols. Those might differ depending on the upstream sample embedding method, as well as on the aimed protein analysis technique.

Please find some examples here:

  • Cryo samples and Western-Blotting (Del Turco et al.)
  • Cryo samples and ESI-MS/MS (Cahill et al.)
  • FFPE samples and LC-MS/MS (Drummond et al.)
  • Cryo samples and LC-MS/MS (Hondius et al.)

6. Protein expression analysis

The following techniques for protein analysis have been successfully tested with proteins gained from LMD:

  • LC-MS/MS
  • ESI-MS/MS
  • SDS-PAGE
  • Western Blotting
  • 2D-DIGE
  • MALDI-TOF
  • SELDI-TOF
  • NMR

Alzheimer protein analysis - Related information

29. April 2019 Laser Microdissection Publication List

This monthly updated reference list demonstrates the major application fields for laser microdissection in life science research.

27. March 2019 Analysis of Tissue Sections Assisted by Fluorescence Laser Microdissection

Lectin microarray (LMA) is a highly sensitive technology used to obtain the global glycomic profiles of endogenous glycoproteins in biological samples…

17. October 2018 Webinar: Live Cell Isolation by Laser Microdissection

Laser microdissection is a tool for the isolation of homogenous cell populations from their native niches in tissues to downstream molecular assays.…

13. September 2018 Nanodroplet Processing Platform for Deep and Quantitative Proteome Profiling of 10–100 Mammalian Cells

Nanoscale or single-cell technologies are critical for biomedical applications. However, current mass spectrometry (MS)-based proteomic approaches…

09. February 2018 Free Webinar On-Demand: How to Conduct Localized Proteomics of Microscopic Regions

In this webinar, you will learn: - How to perform isolated proteomics of microscopic regions of interest - How to use this technique with FFPE…

20. April 2017 Correlation of Phenotype and Genotype – microscopic samples and their biomolecular backgrounds

Seeing is believing, this is what makes microscopes, imaging systems, or box-type imagers so important for researchers in many different fields like…

07. December 2016 MicroRNA Expression Profiling to Identify and Validate Reference Genes for the Relative Quantification of microRNA in Rectal Cancer

Application example for laser microdissection (LMD) - MicroRNAs (miRNAs) play important roles in regulating biological processes at the…

06. June 2016 Workflows & Protocols: Plant Laser Microdissection

During Leica workshops for LMD users in Brazil, hosted by the Federal University of Paraná/UFPR (UFPR) at the Centro de Energia Nuclear na…

04. May 2016 Workflows & Protocols: Laser Microdissection for Pathology and Cancer Research

Tumor development results from mutations in our DNA. For their deeper analysis, cancer researchers have to dissect the relevant tissue areas. Here we…

04. December 2015 Workflows & Protocols: Connecting Microscopy and Molecular Biology in Neuroscience

The main topic during this course was how to apply laser microdissection in neuroscience. Leica specialists demonstrated why laser microdissection is…

Typical fields of research

  • Amyloidosis
  • Cancer
  • Other diseases
  • Drug discovery
  • Plant research

References

  • Drummond et al. Laser Capture Microdissection (2018) pp 319-334
  • Hondius et al. Laser Capture Microdissection (2018) pp 371-383
  • Cahill et al. RapComMS (2018)
  • Drummond et al. ActaNeuropathol 2017

Further information