AMP Core Labs

For planning, note that pricing for Ventana antibodies and reagents increases by approximately 3% once each year.

Cost-based Pricing as of 10/1/23

Cost-based pricing as of 10/1/23

*  Each run also requires at least one positive tissue control, preferably with internal negative controls. Control slides are charged at the same price per slide as above. Additional charges accrue if the protocol requires non-standard ancillary reagents, e.g., amplification ($4.00/slide) or CC2 for antigen retrieval ($1.50/slide).  Note that the costs of Ventana antibodies and ancillary reagents alone predictably increase by 3% per year and these additional costs should be considered when preparing the budget.

As discussed elsewhere, pricing for tests for clinical trials is adjusted to also include costs for a lab administrative component, which will be determined during trial implementation. For most trials this is approximately 20% of the cost of the test.

** Investigators must consult in advance with the Laboratory Manager, Director, or IHC Research technologist. Charges include an initial optimization fee, a run charge based on the number of slides, and charges for Ventana dispensers/prep kits and any non-standard ancillary reagents. The initial optimization/set-up fee is $1200 for a non-Ventana antibody; the time required for optimization is highly variable and success cannot be guaranteed. The optimization fee is reduced to $470 for non-Core Ventana antibodies and waived for non-Core Ventana antibodies with a BJH or DPC Ventana Ultra protocol.

TMA orders are only accepted after prior approval of the project and with a collaborating pathologist to assist in TMA design. Lab functions are limited to the fabrication of previously designed arrays. Because priority must be given to clinical testing, there is no guaranteed turnaround time. Please see the TMA FAQ. The charge for tissue microarrays is per core, regardless of the number of paraffin blocks or cores per block. In addition, an initial technical setup charge and/or hourly pathologist charges will usually apply; an estimate can be provided. The rates for unstained slides and histochemical or immunohistochemical stains are the same as above.

Fixation

The quality of assays performed on fixed and paraffin-embedded tissues is influenced by many pre-analytical factors including the type of fixative, the efficiency and duration of fixation, and the conditions of tissue processing.  For any clinical or research study, it is important to control as many of these variables as possible. For most clinical investigations or clinical trials it is desirable to have biospecimens harvested, fixed, and processed in the same manner as clinical biopsies or excisions.

To this end, the AMP Core Labs has implemented protocols for tissue processing and embedding that closely resemble those used by the pathology labs at Barnes-Jewish Hospital and many (most) other clinical settings. However, initial handling and the initiation of tissue fixation are usually performed by the submitter.

In general, fixation of tissues in the clinical setting relies on standardized commercial preparations of 10% neutral buffered formalin (NBF).   This is a ready-to-use solution of approximately 4% formaldehyde that contains buffer salts and a small amount of methanol.  Buffering is required to minimize time-dependent acidification, which can be deleterious to tissues, while the methanol is required to minimize time dependent polymerization of formaldehyde, which diminishes the efficiency of fixation. Despite these precautions, the fixative has a finite shelf-life, and must be used prior to the indicated expiration date, usually within 2 years of manufacture.

Sources of formalin

The most widely used and reliable source of neutral buffered 10% formalin is Richard-Allan Scientific.  It can be obtained from a large number of different distributors, including Thermo Fisher Scientific. However, several distributors prepackage the fixative in small, polypropylene containers with an inner seal and a 90 ml volume of fixative; this is suitable for most types of small biopsies. Barnes-Jewish Hospital and WU Pathology Services purchase their pre-filled containers from Cardinal Health.

This product provides a safe primary transport container and eliminates the need for staff to handle the fixative. It is important to stock no more NBF than can be used within the estimated shelf-life, to insure reproducible fixation.

Guidelines for formalin fixation of specimens

1. Fixation conditions should be standardized as much as possible. Ideally, tissues should be fixed in formalin for 16-24 hours or more at room temperature to achieve complete penetration and stable crosslinking. Under-fixation is usually more deleterious than over-fixation. Note: the current CAP mandated minimum fixation time for clinical assessment of predictive biomarkers for breast cancer (i.e., ER, PR, and Her-2 receptor expression) is 6 hours, with a maximum time of 72 hours.
2. Tissues should be immersed as soon as possible but within one hour of removal from the body. If absolutely necessary tissues can be briefly placed on a saline moistened gauze sponge; however, even brief placement of a small biopsy on a dry surface can be deleterious.
3. For all clinical studies, including clinical trials, the time of the procedure and the total formalin fixation time must be documented on the requisition and/or specimen container.
4. Tissues should be < 3 mm in thickness to help insure rapid fixation. Although needle or core biopsies do not usually present problem, tissue blocks obtained from larger specimens should be carefully trimmed.
5. After complete fixation in formalin, some research tissues can be safely transferred to 70% ethanol at 4 deg C until processing. As indicated above, under-fixation should be avoided. Note that premature transfer to ethanol can permit reversal of some crosslinks with underprocessing during tissue processing.
   
6. Formalin is the only fixative validated for clinical immunohistochemistry, interphase FISH, or next generation sequencing. For predictive biomarker studies in the clinical setting, alternative fixatives, post-fixation storage in ethanol, and/or decalcification cannot be used without prior validation or appropriate disclaimers on the report.

Precautions when working with formalin

Formalin is a hazardous solution that fixes by gradually penetrating the tissue and crosslinking proteins. Inhalation and physical contact should be minimized. It is important to wear appropriate personal protective equipment and consult with the manufacturer’s MSDS before handling. At a minimum, gloves and eye protection are recommended when working with open containers of formalin.

All specimens submitted for lab services must be properly transported to the lab.

Transport for referred clinical testing at Washington University Medical Center is coordinated by Diagnostic Laboratory Services (DLS).

Clinical research specimens obtained in BJH operating or procedure rooms will usually be delivered by the study coordinator or via the Tissue Repository according to a pre-determined schedule. Transport is coordinated by the study coordinator or designee, not by lab staff.

Regardless of origin, all specimens submitted to the lab must be accompanied by a pre-approved requisition that contains all required identifiers. Note: Clinical trials use a requisition form designed to meet the needs of the trial. However, the requisition must always include the name of the research study or clinical trial, the name of the principal investigator, and the approved unique patient identifiers (usually a pre-assigned patient ID number).

Specimens for clinical testing will only be accepted from approved health care providers. The lab provides no direct to patient testing. Specimens for clinical trials will only be accepted from a registered lab user. Requisitions must always include the full name, title, e-mail address, phone and fax numbers of the submitter(s), i.e. the contact person or physician at the submitting facility.

A “specimen inventory” must be provided; i.e., the specific specimens submitted for analysis must be enumerated and described on the appropriate requisition.The type of fixative and date and time of initial fixation should also be recorded.

Each primary specimen container must have an easily legible, water-proof label affixed to the side of container (not the lid). The label must include: a) required unique patient identifiers, and b) any specimen identifiers if more than one sample is submitted for the same patient. Specimens for clinical trials must also include the study name or number.

Specimens lacking the required information cannot be accessioned, resulting in potential delays. These issues will be brought to the attention of the Lab Manager, Director of Clinical Operations, or Lab Director.

The Lab does not coordinate shipping of research specimens from outside sites but recommends the following:

Packaging, labeling, and shipping are always subject to the relevant International Air Transport Association (IATA) and Department of Transportation (DOT) regulations applicable to diagnostic specimens. The sender should be advised to consult with their institution’s EH&S office or shipping department for more information.

Wet-fixed specimens do not usually require any specific thermal protection. However, paraffin-embeded tissue blocks or unfixed tissues in saline should be shipped with a frozen gel ice-pack during the warmer months.

As an additional precaution against misidentification of a specimen, no more than one patient’s specimen(s) should be shipped per package.

General policies

Any fresh tissues  or tissues in fixative are considered diagnostic specimens for purposes of transport. All specimens require secure packaging, individual specimen labels with required identifiers, and an approved requisition containing all required information.

Transport for referred clinical testing at Washington University Medical Center is coordinated by Diagnostic Laboratory Services (DLS).

Clinical specimens that originate within the Medical Center must use approved specimen containers and be transported by trained pathology transport staff.

The primary container must be a labeled screw top container with a water proof seal. Commercial, pre-filled specimen containers are ideal.  These container(s) should then be placed within a sealable plastic transport bag (e.g., Ziploc) or other approved secondary container.

Research specimens can be hand-delivered to the lab by WU investigators using appropriately labeled primary containers and packaging (see below) and the approved order form.

Clinical or research specimens delivered from outside  the Medical  Center are subject to IATA and DOT regulations and require appropriate, primary, secondary and outer packaging depending on the contents and carrier (see below).  Access Courier is used by BJH and WU Pathology Services.

General guidelines for off-site shipping

The following guidelines are based on CDC recommendations, and are only a guide.  Submitters are responsible for insuring full regulatory compliance by following shipping instructions provided to approved submitters.

Primary packaging

The primary container must be a labeled screw top container with a water proof seal. Commercial, pre-filled specimen containers are ideal. As an additional precaution, the lid should be secured with tape or Parafilm. These container(s) should then be placed within a sealable plastic transport bag (e.g., Ziploc).

For commercial transport, the primary containers should be individually wrapped in sealable plastic bags containing absorbent material; this material will protect the contents from breakage and absorb any leaked contents.

There are volume and weight restrictions for primary containers, but these far exceed the usual specimen shipment.  (The liquid volume or solid weight of primary containers must not exceed 4L or 4 kg; individual primary containers cannot contain more than 1L of liquid.)

Secondary packaging

Use additional and sufficient absorbent material around the primary packaging in the secondary packaging to absorb the entire contents of all primary containers in case of leakage or damage.

Secondary packaging must meet the IATA packaging requirements for diagnostic specimens including 1.2 meter (3.9 feet) drop test procedure.

The secondary container must also be large enough for all markings, labels, and shipping documents (e.g., air waybill).

Leak-proof cold packs must be placed inside insulated packaging, for example Styrofoam box. Outer packaging will be necessary for such packages, as described below.

Outer packaging

An overpack must be used if cold packs are used OR if the secondary packaging is not large enough for all the labels, markings, and documents.

An itemized list of contents must be enclosed just inside the outermost packaging layer, and placed in a sealed plastic bag to protect from moisture.

Labeling of outermost packaging

Each package and the air waybill must be marked with the name, address, and telephone number of the responsible person submitting the package.

For purposes of shipping, both research and clinical specimens should be considered diagnostic specimens. Each package must be labeled as “BIOLOGICAL SUBSTANCE, CATEGORY B” and “UN 3373” in the “Nature and Quantity of Goods” box on the air waybill and outer packaging.

Note: additional packaging and labeling regulations apply in the event dry ice is included or for any international shipments.

All specimens should be addressed to:

AMP Core Labs
Washington University School of Medicine
Department of Pathology & Immunology
425 S. Euclid Ave, Campus Box 8024
4th Floor West Bldg, St Louis, MO 63110

Couriers can deliver specimens directly to Lab 4721 on the 4th floor of the West Building at the South Medical Campus.

The histology and immunohistochemistry areas are organized and equipped to perform laboratory testing for predictive biomarker assays in support of approved clinical trials. Lab management can assist investigators in the early phases of trial development by providing: assay protocols for approved tests, technical and workflow information, and cost estimates.

Predictive biomarkers

The existing test menu includes selected assays of hormone receptor expression and tumor cell proliferation.

Regulatory environment

The Labs are CAP/CLIA-certified and HIPAA compliant. Beyond regulatory compliance, the Labs have a “total test” philosophy that attempts to standardize or define as many variables as possible. This extends from recommendations regarding initial tissue handling and fixation, to standardized processing and embedding, section preparation, and immunostaining.

In vitro diagnostic tests on the Ventana platform

The Labs support selected basic and translational investigations, often employing new immunologic reagents and/or protocols. However, clinical testing is largely restricted to internally verified FDA-approved reagents on the automated Ventana Medical Systems platform.

Accessioning and tracking

Specimens for approved clinical studies can enter the lab in at least 2 ways: as archived paraffin tissue blocks, or as recently harvested tissues in fixative.  The latter tissues are transferred from the Tissue Repository and accessioning in CaTissue. Specimens for trials administered at the Medical Center are accessioned into WU Research CoPath, under the licensing agreement of CoPath Plus with Barnes-Jewish Hospital. However, the anonymized specimens can also be tracked from accessioning through completion of testing via the Lab’s browser-based tracking system, with automatic email notifications at critical steps of the process.

Pathology support

The Core Labs performs predictive biomarker assays and carefully monitors the quality of each assay. However, lab personnel do not interpret or analyze the data. Interpretation rests with the designated pathologist(s) that are co-investigators supported by the grant or contract. The pathologist is usually an attending surgical pathologist in the Division of Anatomic and Molecular Pathology. Results are reported via WU Research CoPath, as for diagnostic specimens.