The Morphology & Imaging module provides technical support in the processing, sectioning, staining, and morphological analysis of ocular cells and tissues at the light and electron microscopic level.

Brief overview of histology services.

Specialized tissue preparation

Most users submit eye tissues for processing that have been chemically fixed by immersion in formaldehyde or similar fixatives.  However, ocular tissue is sometimes best preserved following transcardial perfusion with aldehyde-based fixatives.  This is a more time-consuming procedure but results in the rapid and comprehensive fixation of retina and other vascularized tissues. We can assist with this procedure.  Similarly, rapid freezing (in liquid pentane) followed by freeze substitution in methanol/acetic acid has recently been shown to optimally preserve ocular morphology and enhances staining with certain antibodies (see Sun et al.,2017 Mol Vis 21: 428-442).  We can assist with this procedure also.

Embedding, sectioning, and staining.

The lab performs tissue processing/embedding for routine paraffin samples, frozen sectioning, and glycol methacrylate. The lab offers routine H&E staining and a variety of special stains.  Immunohistochemistry with DAB or fluorescently-labeled secondary antibodies is available and we also offer in situ hybridization (RNAscope) and TUNEL labeling.

Meet the Morphology & Imaging Core team

Steven Bassnett, PhD

Professor, Ophthalmology and Visual Sciences

Below is a list of major equipment than can be used as part of this core

   

Major Equipment

Location

(McMillan)

 1 On-line

Reservation

2 Training

required

Olympus FV1000 MPE confocal microscope

702B

Yes

Yes (Alicia)

Zeiss LSM800 Confocal microscope with Airyscan

702A

Yes

Yes (Alicia)

Olympus BX51 upright Fluorescence microscope

607

Yes

Yes(Alicia)

Olympus BX40 double-headed microscope

607

No

No

Leica Fluorescence Macroscope

622

No

No

Leica LMD6000 laser microdissection system

607

Yes

Yes (Alicia/Belinda)

Bioptigen small animal OCT

606

Yes

Yes (Guang-Yi)

Sakura Tissue-Tek VIP tissue processor

609

n/a

n/a

Shandon Cryotome E cryostat

609

Yes

Yes (Belinda)

Tissue-Tek Paraffin embedding center

609

n/a

n/a

Leica RM2255 microtome

609

n/a

n/a

Leica Ultracut microtome

607

n/a

n/a

In situ hybridization oven

609

Yes

Yes (Guang-Yi)

Image Processing computer with Metamorph software

603

Yes

No

 

1 On-line reservation is available through Facility Online Manager (FOM).

The FOM website address is www.instrumentschedule.com.  Our equipment is listed under WUSTL-VISION. You will need to create a user account in order to access the calendar and reserve time on the instrumentation.  

2 Only individuals trained in the use of this instrumentation will be allowed to reserve time through the FOM website.

For detailed instructions, contact Alicia De Maria (DeMaria@wustl.edu).

Fee Schedule (November 2018)

Confocal microscopy

To offset the cost of the maintenance contracts on the confocal microscopes there is an annual user fee of $4,000 per lab.  This allows unlimited access to the instruments. For occasional users, an hourly fee of $50 is charged.  

Histology pricing

Service

Unit Price ($)

Transcardial perfusion (per mouse)

10

Freeze substitution (freezing in propane, freeze substitution in methanol/acetic acid) (per eye)

10

Tissue processing/embedding – paraffin (per block)

2.50

Tissue processing – frozen (per block)

2.50

Tissue processing – glycol methacrylate (per block)

2.50

Slides – paraffin – unstained (per slide)

0.50

Slides – frozen – unstained (per slide)

0.50

Slides – glycol methacrylate (per slide)

0.50

Histochemical stain (H&E, Toluidine blue, etc) (per slide)

1.00

Immunocytochemistry (with user-provided 1o antibody) per slide

3.00

In situ hybridization (RNAscope) (with user-provided probe) per slide

40.00

TUNEL labeling (to detect apoptotic cells) per slide

15.00