Navigation Links
Innovative Tissue Array Technology for High-Throughput Screening of Gene,,,Expression


Many different tissue types on a convenient single-slide format

Carolyn Thomas
Stratagene

Stratagenes Complete View tissue array slides offer further refinement of tissue array technology through optimized tissue fixation and processing procedures that preserve tissue morphology, antigenicity of proteins, and nucleic acid reactivity. This innovative product line consists of murine (male and female) survey and validation array slides. The Complete View survey array slides represent major organ systems, consisting of approximately 104 cell types mounted in duplicate. The Complete View validation array slides include 12 representative tissues from the survey arrays and are useful to test staining procedures and reagents. We report our results using Complete View array slides for hematoxylin and eosin staining of murine tissue, immunostaining of murine tropomyosin, and labeling RNA of murine salivary gland. These tests confirm that Complete View tissue array slides ensure reliable screening results in a convenient, high-throughput format.

To achieve an understanding of the cellular function of gene products requires localizing gene expression to specific cells and tissues. This task can be accomplished through the use of tissue arrays, a collection of diverse tissue samples mounted on a single slide. Proven successful in examining both normal1,2 and cancerous tissues,3,4 tissue arrays can be treated as a single histological slide during staining, immunohistochemistry, or in situ hybridization. The single-slide format affords a high-throughput method for precisely identifying the specific tissues and cell types that express a gene of interest.

Single-Slide Whole-Animal Surveys

One particularly useful application of tissue arrays is the whole-animal survey, in which all tissues and cell types in an animal species are screened. Traditional sections for whole-animal surveys require approximately 25 to 40 histological slides for complete screening. With each slide requiring preparation and processing that includes staining, labeling, and analysis, such surveys are so demanding of time, labor, and reagents that they are impractical in most research environments. In contrast, a single Complete View tissue array can contain all the major tissues and cell types of an animal. Hence, tissue array technology makes it possible to screen numerous tissues in a single experiment (Figure 1).

Fig.1

Stratagene has improved tissue array technology by creating the Complete View survey and validation array slides. In this product line, optimized tissue fixation preserves morphological integrity, antigenicity of proteins, and nucleic acid reactivity. On each Complete View array slide, tissues are conveniently configured for easy identification with the Complete View target locator, which positions each tissue within the array by column and row. The availability of two categories of array slides provides further convenience. The Complete View survey array slides represent major organ systems that consist of approximately 104 cell types mounted in duplicate (Table 1). The Complete View validation array slides include representative tissues from the survey arrays and are useful to test staining procedures and reagents.

Slides

Tissue specimens

Murine Validation Array,
Male C57BL/6J or
Female C57BL/6J

12 unique tissue types represented from the following groups:

  • connective tissue
  • skeletal tissue
  • nervous tissue
  • lymphatic tissue
  • integument
  • alimentary system
  • respiratory system
  • urinary system
  • glandular tissue

Each representative tissue has dual representation per slide.

(24 tissue spots per slide + 1 positive and 1 negative control spot)

Murine Survey Array,
Male C57BL/6J or
Female C57BL/6J

In excess of 40 tissue types representing greater than 100 cell types from the following groups:

  • connective tissue
  • skeletal tissue
  • nervous tissue
  • lymphatic tissue
  • integument
  • alimentary system
  • respiratory system
  • urinary system
  • glandular tissue
  • vascular system
  • reproductive system

Preserved Tissue Integrity

We assessed the morphological integrity of tissues and the antigenicity of proteins by assaying the Complete View array slides in hematoxylin and eosin staining and immunohistochemical analysis, respectively. Figure 2 shows multiple tissues stained within the Complete View male mouse survey array slide. To test antigenicity, we used the Complete View male mouse validation array slide as described (Methods) with antitropomyosin as the primary antibody and goat anti-rabbit FITC as the secondary antibody. The result of this immunological screening assay demonstrates the actin-binding protein within the muscle tissue (Figure 3).

Fig.2

Fig.3

Since many applications of tissue array technology rely on nucleic acid assays, we used the Complete View male mouse validation array slides for an in situ hybridization experiment (Figure 4). One of two slides was treated with RNase prior to hybridization of both slides with a fluorescently labeled poly(dT) probe. Salivary gland tissue on the slide without RNase digestion clearly shows the presence of mRNA . We used standard conditions for slide preparation, prehybridization, and hybridization reactions (Methods).

Fig.4

Conclusions

The Complete View tissue array slide kits permit wide- spread use of tissue array technology. With the single-slide format and convenient configuration of tissues, these innovative array slides simplify screening procedures so specialized expertise in histology is not required. We demonstrated their use to localize expression to specific tissues and cells, and our data confirm that these optimized array slides maintain the morphological integrity of tissues. By immunostaining of the tropomyosin protein and fluorescent labeling of RNA, we verified that the tissues affixed to the Complete View slides retain antigenicity of proteins and reactivity of nucleic acids, respectively. We are expanding this product line to include human fetal validation and survey arrays.

Methods

In Situ Hybridization: Paraffin was removed from the array tissue by treating the slides as described in the product instructions. The two consecutive slides were rehydrated and washed in PBS, immersed in 0.1 M glycine, and washed three times in PBS. Then, the slides were incubated with proteinase K and one was subsequently treated with RNase. To stop the reactions, slides were immersed in 4% paraformaldehyde in PBS followed by a PBS rinse. The slides were then incubated with acetic anhydride in triethanolamine and prehybridized in formamide (50% v/v) in 4X SSC at 37C for 30 minutes, then hybridized with 20 ml of poly d(T)-FITC incubated at 42C for 12 to 18 hours. This incubation was followed by a series of washes with 4X SSC. Each slide was examined with an Olympus BX60 fluorescent microscope.

Immunohistochemistry: Paraffin was removed from the array tissue by treating the slide as described in the product instructions. The slide was rehydrated, then incubated in PBS for 5 minutes and placed in a humidity chamber. The diluted primary antibody was added to the slide and incubated for 30 to 60 minutes at 37C. After washing with PBS, the diluted secondary antibody was added to the slide and incubated for 30 to 60 minutes at 37C. Following two additional PBS washes, a coverglass was placed over the slide, and the slide was examined using an Olympus BX60 fluorescent microscope.


REFERENCES
  1. Battifora, H. (1986) Lab. Invest. 55(2): 244-248.
  2. Battifora, H. and Mehta, P. (1990) Lab. Invest. 63(5): 722-724.
  3. Kononen, J., et al. (1998). Nature Med. 4(7): 844-847.
  4. Bubendorf, L., et al. (1999) Cancer Res. 59: 803-806.

'"/>

Source:


Page: All 1 2 3 4 5

Related biology technology :

1. Detecting Gastrin-Releasing Peptide Receptor by in situ PCR on Archived Tissue
2. Simple Isolation of RNA from Tissue and Cultured Cells
3. Tissue Specificity for Mutation Parallels Tissue Specificity for Cancer
4. Hands-free Tissue Disruption No Homogenization Needed
5. Recover High Yields of Total Nucleic Acid from Formalin-fixed, Paraffin-embedded (FFPE) Tissue
6. RNase Activity in Mouse Tissue: Classification, Hierarchy, and Methods for Control
7. Stabilizing RNA in Fresh and Frozen Tissues for RNA Isolation
8. Genomic DNA Preparation from RNAlater Preserved Tissues
9. DNA Isolation From 300 to 450 mg Dried or 600 to 1,200 mg Fresh Leaf Tissue
10. DNA Isolation From 50 to 20 mg of Paraffin-Embedded Tissue
11. Genomic DNA Isolation From 510 mg Marine Invertebrate Tissue
Post Your Comments:
(Date:8/19/2014)... (PRWEB) August 19, 2014 ... to raise $200,000 to fund a corporate lab ... , This development is a critical component ... of care to the implementation of personalized medicine ... of practicing medicine. , Indiegogo contributions will support ...
(Date:8/19/2014)... GLEN BURNIE, Md. , Aug. 19, 2014 /PRNewswire/ ... the clinical research industry, recently announced the formation of ... is pleased to announce the addition of Dr. ... Photo - http://photos.prnewswire.com/prnh/20140818/136957 ... insight across a variety of therapeutic areas and set ...
(Date:8/18/2014)... Dallas, Texas (PRWEB) August 19, 2014 ... China Linalool Industryā€¯ is a professional and in-depth ... report provides basic Linalool information, including its ... well as industry overview. This research covers the ... well as global industry analysis covering macroeconomic environment ...
(Date:8/18/2014)... with unprecedented efficiency and data storage may ... built-in electric polarizations to read and write ... popular data-driven technology. But ferroelectrics must first ... a curious habit of "forgetting" stored data. ... Department of Energy,s Brookhaven National Laboratory have ...
Breaking Biology Technology:Cellgen Diagnostics Launches Indiegogo Crowdfunding Campaign for DNA Medical Testing Device 2Cellgen Diagnostics Launches Indiegogo Crowdfunding Campaign for DNA Medical Testing Device 3CSSi Appoints Dr. William E. Gannon, Jr. to Newly Formed Medical and Clinical Advisory Board 2CSSi Appoints Dr. William E. Gannon, Jr. to Newly Formed Medical and Clinical Advisory Board 3Linalool Industry & Diphenylamine Market (Global, China) 2014 Analysis Now Available at DeepResearchReports.com 2Linalool Industry & Diphenylamine Market (Global, China) 2014 Analysis Now Available at DeepResearchReports.com 3Linalool Industry & Diphenylamine Market (Global, China) 2014 Analysis Now Available at DeepResearchReports.com 4Promising ferroelectric materials suffer from unexpected electric polarizations 2Promising ferroelectric materials suffer from unexpected electric polarizations 3Promising ferroelectric materials suffer from unexpected electric polarizations 4
... Calif., Nov. 26 NeurogesX, Inc.,(Nasdaq: NGSX ... management therapies, announced today that it is scheduled ... Lazard Capital Markets Healthcare,Conference in New York City. ... presenting and Stephen Ghiglieri, Chief Financial Officer,will be ...
... Nov. 26 Archer Daniels Midland,Company (NYSE: ... vice,president, has elected to retire by year end., ... special assignment to advance the,Company,s business strategy for ... to the Company,s strategic planning group,under the direction ...
... (Amex:,SGN - News) has received FDA 510(k) clearance ... ambulatory event recorder throughout the United States.,This device, ... HeartLife,Tempo Card, is a direct-to-consumer non-prescription flexible credit ... as an early-detection,device by patients who desire to ...
Cached Biology Technology:NeurogesX to Present at Lazard Capital Markets Healthcare Conference 2NeurogesX to Present at Lazard Capital Markets Healthcare Conference 3ADM Executive Vice President Bill Camp to Retire 2Signalife Receives FDA Clearance for OTC Non-Prescription Event Recorder Device 2Signalife Receives FDA Clearance for OTC Non-Prescription Event Recorder Device 3
(Date:8/19/2014)... (Boston) Some infants born with neonatal abstinence ... a more difficult time going through withdrawal than ... understood. While genetic and epigenetic (when genes are ... identified as potential factors, researchers at Boston University ... (BMC) conducted a first of its kind study ...
(Date:8/19/2014)... had a glass of fizzy soda knows that bubbles ... a finding with wide industrial applications, Princeton researchers have ... into the liquid as well. , "It is well ... were surprised, and fascinated, to discover that when we ... tiny oil droplets into the water," said Howard Stone, ...
(Date:8/19/2014)... new study that analyzed concentrations of African dust ... in winter and spring. These research findings offer ... air quality impacts of African dust, including the ... to occur from increased African dust emissions in ... Miami (UM) Rosenstiel School of Marine and Atmospheric ...
Breaking Biology News(10 mins):In-utero methadone, Subutex exposure could alter gene expression, cause severe Neonatal Abstience Syndrome 2Bubbling down: Discovery suggests surprising uses for common bubbles 2Bubbling down: Discovery suggests surprising uses for common bubbles 3Study of African dust transport to South America reveals air quality impacts 2
... reaction (PCR) is a common technique used to amplify, or ... in genetic analyses for everything from medicine to forensics. ... and sequencing genes, and its applications are endless. However, ... the University of Southern Mississippi discovered that the use of ...
... an embryonic stem cell is in the first stage of ... type of cell in the body, a state scientists call ... a way to easily distinguish undifferentiated embryonic stem cells from ... published in the American Institute of Physics, (AIP) journal ...
... apply for press credentials for the American Chemical Society,s ... conferences of the New Year will be held April 7-11, ... than 13,000 scientists and others expected to gather in the ... span science,s horizons from astronomy to zoology ― the ...
Cached Biology News:Improving DNA amplification from problematic plants 2Press registration opens for 2013 meeting of world's largest scientific society 2
Animal Origin Free...
IHC Detection Kit (Broad Spectrum AEC)....
...
... Antibody Modification Services ... including antibody labeling and antibody fragmentation. ... provides signals for visualization and quantitation ... assays, it is preferable to use ...
Biology Products: