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DERC Mouse Phenotyping | The Histopathology Laboratory | Body Composition |
Multiplex Protein Assays (MPAs) | Whole Body Metabolic Assessment
Histopathology
The Histopathology Laboratory of the Mouse Phenotyping Core provides a wide spectrum of sample preparation services. Originally processing only pancreatic tissue, the lab is now able to embed, section, and stain almost all tissues relevant to the study of diabetes (pancreatic, adipose, liver, central nervous system, skeletal muscle, cardiovascular, gonadal, and kidney tissues).
The laboratory has also expanded its immunohistochemical techniques to a wide range of tissues. Currently active protocols exist for mouse antigens listed in Table 1.
Pancreas related/ relevant antigens
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Adipose Tissue related/relevant cell markers |
CNS-Related Antigens
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Cell Lineage Markers |
Insulin Signaling Antigens |
Hepatic Antigens |
Inflammation |
Growth Proliferation/ Apoptosis
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Insulin
C-peptide Glucagon Pdx1
Nkx2.2 Nkx6.1 NeuroD MafA, B Ngn3
Runx1 Vimentin Foxo1/3 Lisch-like HNF-3B Nestin
Isl-1
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Perilipin
ADRP
F4/80
TNF a Mahoganoid beta-Catenin Foxo1/3
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Robo1
Slit1
Slit2
cFos Mahogany
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N-cadherin Vimentin
E-cadherin Smooth muscle beta-Actin Desmin Cytokeratin 1 Cytokeratin19 Pan-Cytokeratin Amylase
GFP
eGFP
LacZ Hemagglutin
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IRS1
IRS2
PDK1
PTEN
S6 kinase pS6 kinase AKT
pAKT Foxo1/3
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Notch3 Foxo1/3 Glut2 IRS1 IRS2
PDK1
PTEN
S6 kinase pS6 kinase AKT
pAKT
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F4/80
MMP1
MMP3
MMP8 MMP12 MMP13
CD3e
CD45.1 CD45.2
TNFa
Ly6g
CD68
myelo-peroxidase CD45R CD11b ICAM1
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Ki67
cMyc
activated- Caspase3 PNCA cFos
BRDU
TUNEL
p-histone-2Hb EGFR
K-ras
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Table 1. Mouse antibodies currently available for immunohistochemistry
Additionally, the lab has been able to add immunofluorescence-based services, enabling investigators to study multiple antigens in a single section. Protocols have been established for antigens expressed in most of the metabolically relevant tissues including: Insulin, Glucagon, Foxo1, F4/80, CD45.2, ICAM1, CD3e, eGFP, LacZ, c-Myc, and Caspase3.
Antibodies for mouse IHC
Location, Equipment and Staff:
The histopathology laboratory is located in a dedicated wet laboratory adjacent to the laboratories of DERC investigators Rudy Leibel and Arg Efstratiadis. It is fully equipped with two microtomes, a cryostat , automated embedding system, automated staining system (DAKO ) and a light microscope. The staff of the histopathology laboratory is supervised by Qiong Li, who is supported by two senior technicians. Matthias Szabolcs, MD, provides general consultation and meets with the staff to review work and troubleshoot problems on a weekly basis.
Procedure for using the Histopathology Laboratory:
DERC investigators provide fixed samples to the Histopathology Lab using fixation protocols provided by the Core. Samples are registered by the Lab and processed. Completed samples are then returned to the investigator for analysis, following which an email is sent to assess quality. Investigators purchase and provide primary antibodies or specialty staining materials. A fee is charged for service as listed on the DERC website and in the table below. If perfusion fixation is required, the investigator schedules a time with the Laboratory staff and brings mice to the laboratory where they are sacrificed by CO2 asphyxiation immediately prior to perfusion.
While Non-DERC users do have access to the services provided by the histopathology laboratory, DERC users get first priority and pay a discounted price.
PRICE LIST |
DERC
Member |
Non-Member |
Histopathology |
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Gross room preparation of tissue per cassette |
$ 2.00 |
$ 3.00 |
Machine processing and embedding |
$ 5.00 |
$ 7.50 |
H&E staining per slide |
$ 2.00 |
$ 3.00 |
Cutting one section of blanks starting with a block |
$ 1.00 |
$ 1.50 |
Cutting one section of immuno blanks starting with a block |
$ 1.25 |
$ 1.88 |
Re-embedding one paraffin block |
$ 2.00 |
$ 3.00 |
Histochemical Staining |
$ 10.00 |
$ 15.00 |
Cutting frozen sections (1-2 sections) |
$ 5.00 |
$ 7.50 |
Cutting frozen sections (3rd and subsequent) |
$ 2.00 |
$ 3.00 |
Preparing block for frozen sectioning |
$ 5.00 |
$ 7.50 |
IHC staining per slide (autostainer) |
$ 15.00 |
$ 22.50 |
IHC staining per slide (manual) |
$ 20.00 |
$ 30.00 |
FIHC staining per slide (manual) |
$ 30.00 |
$ 45.00 |
Custom procedures, e.g. perfusion-fixation and microscopy services (hourly rate) |
$ 60.00 |
$ 90.00 |
Quality Control:
The laboratory has three methods of quality control. First, an assessment is sent and completed by every user of the lab. These are quick and simple, and are reviewed by the lab staff. Second, an internal review is performed weekly, during which sample slides from the previous week are reviewed. Third, every six months images representative of the samples processed are sent to expert consultants to assess their quality. Their comments are then reviewed with the Histopathology Lab staff and the Core Director.
Contact Number - 212-851-5312
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