Navigation Links
Laser Capture Microdissection of Cells Labeled with Enhanced Green Fluorescent Protein

Vasker Bhattacherjee, Partha Mukhopadhyay, Saurabh Singh, Robert M. Greene and M. Michele Pisano, University of Louisville Birth Defects Center, Department of Molecular, Cellular and Craniofacial Biology, Louisville, KY 40202


Abstract
This application note describes a Laser Capture Microdissection (LCM) method for the isolation of cells expressing enhanced green fluorescent protein (EGFP) from frozen sections of embryonic tissue. The application of this methodology was used to facilitate retrieval of RNA from mouse embryonic neural crest cells that could not be distinguished by morphology, histological stains or labeling by fluorescent antibodies in a heterogeneous tissue. Specificity of the LCM of the neural crest cells was confirmed by quantitative real-time PCR of neural crest gene markers.


Introduction
The emergent technology of Laser Capture Microdissection (LCM; Arcturus; Emmert-Buck et al., 1996; Simone et al., 1998) is used to isolate specific cell types from tissue sections for downstream molecular analyses. In traditional applications, LCM is used to isolate cells that are distinguished by morphology, histological stains, or labeling by fluorescent antibodies from a heterogeneous population of cells in a tissue section. In this Application Note, an adaptation of this technique for the microdissection of cells genetically labeled with enhanced green fluorescent protein is described (EGFP; Bhattacherjee et al., 2004). EGFP is a recombinant protein originally derived from the jellyfish, Aequoria victoria, which has the advantage over other reporters of not requiring cell/tissue fixation for visualization. EGFP fluoresces green when excited by light of 488 nm and may be visualized by epifluorescence microscopy using standard fluorescein isothiocyanate filters (Chalfie et al., 1994) under wavelengths that are not deleterious to living tissues. Since EGFP consists of ~200 amin
'"/>

Source:


Page: All 1 2 3 4 5

Related biology technology :

1. Isolate and Analyze Total RNA from Cells Harvested by Laser Capture Microdissection
2. Laser microdissection and nucleic acid purification - a Leica - QIAGEN cooperation
3. Working With Laser Capture Microdissection Samples
4. Laser Capture Microdissection (LCM) for the Analysis of Macrophage Gene Expression from Atherosclerotic Lesions
5. Laser Capture Microdissection (LCM) and Expression Profiling of Long-Range Projection Neurons
6. Differential Gene Expression Analysis of Pure Cell Populations from Frozen UterineTissue Using Laser Capture Microdissection (LCM) and cDNA Microarrays
7. Laser Capture Microdissection of muscle fiber populations and expression analysis by RT-PCR
8. An Epitope Tagging Vector for Gene Expression in Mammalian Cells
9. Versatile Vectors for Ponasterone A- Inducible Control of Gene Expression in Mammalian Cells
10. Cell Proliferation Kit Outperforms the Competition in the Range of Cells Typically Counted
11. XL10-Gold Cells Supplied in New Single-Reaction Format
Post Your Comments:
(Date:5/19/2013)... Switzerland’s MS Tûranor PlanetSolar , the ... record for a transatlantic crossing by a solar electric ... 25, 2013, the solar powered boat sailed 2,867miles (5,310 ... of 5.3 knots before reaching Marigot, St. Martin, in ... first-of-its-kind catamaran completed this year’s passage across the Atlantic ...
(Date:5/17/2013)... 17, 2013 Oven Industries Inc. announces ... 5R6-900 temperature controller has many user-friendly benefits. Contained all ... the wall as a self-contained temperature control ... detail makes the device unique, as well as highly ... also be used universally, which allows the user to ...
(Date:5/17/2013)... Cardiff, UK (PRWEB) May 17, 2013 ... by Photonics21 at the association’s recent annual ... others including EC Vice-President Neelie Kroes, who called the ... between sectors, and a stimulant to European innovation.” , ... the structure of the new Photonics PPP at its ...
(Date:5/17/2013)... 17, 2013  Yongye International, Inc. (NASDAQ: ... manufacturer, and distributor of crop nutrient products in ... announced that, on May 16, 2013, the special committee ... "Board of Directors") was provided a letter (the "Letter") ... ) Limited ("Abax") to Full Alliance International Limited ...
Breaking Biology Technology:MS Tûranor PlanetSolar Sets World Speed Record for Transatlantic Crossing by Solar Electric Vessel 2MS Tûranor PlanetSolar Sets World Speed Record for Transatlantic Crossing by Solar Electric Vessel 3MS Tûranor PlanetSolar Sets World Speed Record for Transatlantic Crossing by Solar Electric Vessel 4MS Tûranor PlanetSolar Sets World Speed Record for Transatlantic Crossing by Solar Electric Vessel 5MS Tûranor PlanetSolar Sets World Speed Record for Transatlantic Crossing by Solar Electric Vessel 6New Temperature Control Products Announced by Oven Industries Inc. 2SPIE Joins in Praise for ‘Huge Step Forward’ for Innovation R&D Enabled by Photonics21 Actions 2SPIE Joins in Praise for ‘Huge Step Forward’ for Innovation R&D Enabled by Photonics21 Actions 3Yongye International Provides Update on Status of Proposed Go Private Offer 2Yongye International Provides Update on Status of Proposed Go Private Offer 3Yongye International Provides Update on Status of Proposed Go Private Offer 4
... ... with regulatory agencies in Latin America regarding approvals of Stemedica’s allogeneic adult stem cell products ... the United States. , ... (PRWEB) June 24, 2010 -- Stemedica Cell Technologies, Inc., a leader in ...
... Spain , June 24 Spanish biotechnology company ... has granted Grifols exclusive world-wide distribution rights for its BLOODCHIP® ... blood donors to assure the compatibility of blood for transfusion ... , , ...
... Vt ., June 22 As previously announced on October ... of the Del-ID system to an Ontario company, QTECH SYSTEMS INC. ... announce leadership changes and appointments to QTECH,s Management Team and the appointment of ... . , , , ...
Cached Biology Technology:Stemedica Announces the Appointment of Dr. Csar Amescua Garcia as Medical and Regulatory Affairs Director – Latin America 2Progenika Biopharma Grants Grifols World-Wide Distribution Rights for the BLOODCHIP® Genotyping Test for Blood Donors 2Progenika Biopharma Grants Grifols World-Wide Distribution Rights for the BLOODCHIP® Genotyping Test for Blood Donors 3delSECUR CORPORATION Announces Organizational Update as QTECH SYSTEMS INC. Expands Leadership Team 2delSECUR CORPORATION Announces Organizational Update as QTECH SYSTEMS INC. Expands Leadership Team 3
(Date:5/17/2013)... of a group of proteins in the brain responsible ... new target that could increase cell survival. , The ... Biochemistry and published in the EMBO journal ... to new therapies for stroke and other brain diseases. ... team which identified a protein, known as SUMO, responsible ...
(Date:5/17/2013)... human-caused climate change may have little impact on ... of recent studies that predict their widespread extinction ... which appear in the journal Global Change ... a creature thought to be doomed: http://onlinelibrary.wiley.com/doi/10.1111/gcb.12253/abstract ... especially forest lizards, will be hard hit by ...
(Date:5/16/2013)... 2013) Illustrating a commitment to the support ... Association (AGA) Research Foundation has announced the inaugural ... Award recipients. Supported by the National Institute of ... new award helps underrepresented minority students to further ... research. , "By establishing this new award, AGA ...
Breaking Biology News(10 mins):SUMO wrestling cells reveal new protective mechanism target for stroke 2Climate change may have little impact on tropical lizards 2Underrepresented minority students receive fellowships in digestive disease and nutrition research 2
... and center of today,s political debates, the other indicators of ... in the public conscience. Featured as part of the ... newsstands today, U of M professor Jon Foley makes the ... all environmental processes that contribute to the Earth,s health. In ...
... University] Protein phosphorylation is a process by ... to another. It is a crucial function for ... cellular process, including metabolism, gene transcription, cell-cycle progression, ... its importance in biology, scientists have wanted to ...
... team of researchers, led by a UC Davis veterinary endocrinologist, ... in rats that is similar to bariatric surgery in humans ... also have identified biochemical changes caused by the surgeries that ... study, published online in the journal Gastroenterology , should ...
Cached Biology News:Key enzyme discovered to be master regulator in protein-protein reactions 2New study may explain how weight-loss surgery reverses type 2 diabetes 2New study may explain how weight-loss surgery reverses type 2 diabetes 3New study may explain how weight-loss surgery reverses type 2 diabetes 4
WD-repeat protein 4...
Rabbit polyclonal to MDM2 (phospho S185) ( Abpromise for all tested applications). entrezGeneID: 4193 SwissProtID: Q00987...
... Cultured human omental preadipocytes ... but otherwise healthy donors. ... of diabetic cells. Please ... Support media also available. ...
ASCIZ...
Biology Products: