Cancer Awareness – Cervical Cancer Facts

Approximately 13,000 women in the United States are diagnosed with cervical cancer annually, leading to an annual estimated 4,000 deaths.1 Cervical cancers are comparatively rare in the United States, with about 0.8% of new cancer cases classified as cervical, contributing approximately 0.7% of cancer deaths yearly.1 Worldwide, cervical cancer is the fourth most frequently diagnosed cancer and the fourth leading cause of cancer death, with an annual estimated 527,600 cases and 265,700 deaths.2

Nearly all cervical cancers are caused by high-risk (oncogenic) human papilloma virus (HPV).3,4 HPV subtypes are broadly classified as high-risk (leading to cervical malignant transformation) or low-risk (leading to benign lesion development).5 The high-risk HPV subtypes, HPV 16 and HPV 18, are the most common subtypes found in cervical tumors. Together, HPV 16 and HPV 18 are the causative agents for greater than 70% of invasive cervical cancers worldwide.5,6 It has been estimated that approximately 10% of the worldwide population have a cervical HPV infection at any given time.7 In spite of this high infection rate, nearly 90% of such infections are naturally cleared by the body within a year or two, and only women with persistent high-risk HPV subtype infections are at risk of progression to cervical cancer.8,9 The slow progression of the disease combined with the availability of screening and HPV vaccination is why many experts consider cervical cancer to be nearly completely preventable.10 Invasive cervical cancer can often be successfully treated if detected at an early stage.

As part of our promise of “Fighting Cancer, One Slide at a Time,” Biocare Medical is proud to offer key high-quality immunohistochemistry (IHC) and in situ hybridization (ISH) reagents that aid in early and accurate classification of cervical tumors and presence of HPV.

Key Antibodies for Cervical Cancer

Product Name Clone Catalog Number
HPV-16* CAMVIR-1 CM 186
HPV Cocktail Broad Spectrum* BPV-1/1H8 + CAMVIR-11 CM 177; PM 177
CA 125 OC125 CM 101; PM 101
Carcinoembryonic Antigen (CEA {M}) COL-1 CM 058; PM 058
p63 4A4 CM 163; PM 163; IP 163; OAI 163; VP 163
CD117/c-kit EP10 CME 296; PME 296; IP 296; OAI 296
MCM2 + TOP2A OT18A11 + UMAB146 API 3181
Topoisomerase II alpha O31 ACI 3045; API 3045
VEGF* EP1176Y CME 356

*Research Use Only (RUO)

Key probes for detecting HPV

Product Name HPV Target Subtype (Classification) Catalog Number
HPV Type 16 Probe (Digoxigenin)** HPV 16 (High-Risk) BRA 4047
HPV Type 18 Probe (Digoxigenin)** HPV 18 (High-Risk) BRA 4048
HPV Type 31 Probe (Digoxigenin)** HPV 31 (High-Risk) BRA 4051
HPV Type 51 Probe (Digoxigenin)** HPV 51 (High-Risk) BRA 4052
HPV Type 6 Probe (Digoxigenin)** HPV 6 (High-Risk) BRA 4045
HPV Type 11 Probe (Digoxigenin) HPV 11 (High-Risk)** BRA 4046

**Analyte Specific Reagents (ASR) for clinical Laboratories to use in developing laboratory developed tests.

Clinical References:

1. Siegel RL, Miller KD, Jemal A. CA Cancer J Clin. 2017 Jan;67(1):7-30.
2. Torre LA, et al. Cancer Epidemiol Biomarkers Prev. 2017 Apr;26(4):444-457.
5. de Sanjose S, et al. Lancet Oncol. 2010 Nov;11(11):1048-56.
6. Li N, et al.. Int J Cancer 2011;128:927–35.
7. de Sanjose S, et al. Lancet Infect Dis 2007;7:453–9.
8. Shvetsov YB, et al. Clin Infect Dis 2009;48:536–46.
9. Moscicki AB, et al. Vaccine 2012;30Suppl 5:F24–33.
10. Committee on Adolescent Health Care Immunization Expert Work Group. Obstet Gynecol 2015;126:e38–43.

Heat-Induced Epitope Retrieval


Formalin has been used since 1893 as the standard fixative for tissue processing in histopathology.1 Due to formalin’s superior preservation of morphological detail, criteria for pathological diagnoses have been established through the observation of formalin-fixed, paraffin-embedded (FFPE) tissue sections stained with hematoxylin and eosin. Although many other fixatives exist, none have supplanted formalin in general use. Formalin is a cross-linking type of fixative, forming methylene bridges between proteins within the tissue.2,3 This cross-linking reaction adversely alters the structure of tissue proteins, resulting in loss of antigenicity.4 One way to restore antigenicity is through the use of heat-induced epitope retrieval (HIER) prior to immunohistochemistry (IHC).

The complexity of the IHC protocol demands a properly-trained, highly-skilled staff to achieve the most accurate, consistent diagnoses. As the enhanced diagnostic utility of IHC has been realized, an increased demand has been placed on the histology laboratory. Automation, and the concomitant standardization and reduction of variability, allows laboratories to achieve the quality, reproducibility, and speed necessary to meet the increased IHC demand.5


HIER, also known as Antigen Retrieval (AR), is based upon biochemical studies suggesting that the cross-linking reaction between protein and formalin may be reversed by high-temperature heating or strong alkaline hydrolysis.2,3,6 The routine use of HIER prior to IHC has been shown to minimize inconsistency and standardize staining.7-8 The most critical aspects that influence the quality of HIER results: 1) Heating temperature; 2) Heating time; and 3) The pH of the retrieval solution.

HIER is the process of heating the slide-mounted specimen material in an antigen retrieval solution, followed by a cooling-off period. There is no universal antigen retrieval solution available for restoring all antigens.9 The most commonly used antigen retrieval solutions are citrate-based, tris-based, or EDTA-based solutions.9-10 The pH of the HIER solution influences the immunohistochemistry staining intensity significantly and is critical to the effectiveness of HIER for the specific antigen tested.11 HIER at the appropriate pH allows the antigen to regain its natural shape. Some antigens retrieve better in a lower pH solution, such as a citrate buffer, while others will retrieve better in a higher pH solution, such as a tris buffer.9,11 The composition and the pH of retrieval buffers are critical for optimal retrieval and subsequent staining.

One mechanism to explain how HIER works is by hydrolysis of the methylene bridge cross-links.9,11,12,13,14 Breaking the formalin-induced methylene cross-links in the presence of the appropriate pH solution allows the fixed protein to undergo a series of conformational changes to restore, or partially restore, its native structure. The conformational change allows for better access for the antibody to bind to the antigen. The entire process is being driven by thermal energy from the heat source.11 It is hypothesized that the application of heat accelerates breakage of the methylene cross-links. Subsequently, the methylene cross-links get hydrolyzed when that energy threshold has been reached. The amount of energy added during HIER is determined by heating temperature as well as heating time. There is an inverse relationship between heating temperature and heating time: the higher the temperature, the shorter the heating time needed to attain optimal results. Heating at a higher temperature for a shorter duration yields better staining results than heating at a lower temperature for a longer time.9

The source of heat can be from any number of laboratory equipment, including: pressure cookers, steamers, water baths, microwave ovens, and fully automated IHC stainers. Using a laboratory pressure cooker eliminates the irregular heating and temperature variation/hot spots inherent to steamers and microwave ovens.9,10,13 Ultimately, the introduction of automated IHC staining platforms and online heat retrieval techniques solved much of the inconsistency problems encountered with the majority of other heat retrieval devices. Online HIER processes are more reproducible, less damaging to tissue sections, and save a great deal of time. One major attribute of automation is standardization of protocol and reducing variability from user to user with manual HIER and IHC staining processes.

Clinical References:

1. Fox CH, et al. J Histochem Cytochem. 1985 Aug;33(8):845-53.
2. Fraenkel-Conrat H, Olcott HS. J Biol Chem. 1948 Jul;174(3):827-43.
3. Fraenkel-Conrat H, Olcott HS. J Am Chem Soc. 1948 Aug;70(8):2673-84.
4. Montero C. J Histochem Cytochem. 1991 Jun;39(6):741-8.
5. Prichard JW. Arch Pathol Lab Med. 2014 Dec;138(12):1578-82.
6. Shi S-R, Key ME, Kalra KL. J Histochem Cytochem. 2003 Jan;51(1):1-4.
7. Shi S-R, Cote RJ, Taylor CR. J Histotechnol. 1999 Sep;22(3):177-92.
8. Taylor CR, et al. Biotech Histochem. 1996 Jan;71(5):263-70.
9. Ramos-Vara JA, Miller MA. Vet Pathol. 2014 Jan;51(1):42-87.
10. Tacha D, Teixeira M. J Histotechnol. 2002 Dec;25(4):237-42.
11. Shi S-R, Cote RJ, Taylor CR. J Histochem Cytochem. 2001 Aug;49(8):931-7.
12. Guo T, et al. J Histochem Cytochem. 2007 Jul;55(7):763-72.
13. Van Hecke D. J Histotechnol. 2002 Mar;25(1):45-54.
14. Gown AM. Am J Clin Pathol. 2004 Feb;121(2):172-4.

Cancer Awareness – Early Detection and Tumors of Unknown Origin

Biocare Medical takes great pride in our promise of “Fighting Cancer, One Slide at a Time.” Our focus in the fight against cancer is realized through our high-quality immunohistochemistry (IHC) and in situ hybridization (ISH) reagents and instrumentation. Specifically, our sensitive and specific antibodies, molecular probes, and detection chemistries support early and accurate classification of many cancer types, including tumors of unknown origin. Early detection of cancer significantly increases the chances for successful treatment and can effectively reduce the associated mortality, while accurate diagnosis of tumors of unknown origin allows for determination of the best treatment option.

Early Diagnosis

Early diagnosis for the nine most common cancers (bladder, colon, breast, cervical, ovarian, malignant melanoma, lung, prostate, and testicular cancers), significantly increased survival rates1. When the cancer was diagnosed at stage one or two, 80% of patients survived for at least 10 years.

The appropriate immunohistochemical panels, as well as ISH and other molecular testing, are a critical component in the systematic algorithm of specific, accurate diagnosis2. In the past decade, the clinical diagnostic precision of organ- and tumor-specific immunomarkers, along with the clinical validation of effective IHC panels, has improved significantly.

Tumor of Unknown Origin

When the lineage of a tumor can be identified through standard morphological means, organ- or tumor-specific immunomarkers would be utilized to determine an accurate diagnosis. If the tumor lineage cannot be easily identified via morphological analysis, IHC plays a critical role in the determination of the site of primary origin2,3. Often, the first diagnostic IHC panel will include antibodies directed against lymphoid antigens (Leukocyte Common Antigen [LCA] Cocktail), epithelial antigens (Pan Cytokeratin [AE1/AE3]), melanocytic antigens (S100 protein, SOX10), and sarcoma antigens (Muscle Specific Actin).

Mainline Screening Immunomarkers for Tumors of Unknown Origin

Lymphoma Carcinoma Melanoma Sarcoma
Leukocyte Common Antigen (LCA) Cocktail Pan Cytokeratin [AE1/AE3] S100 Protein Cocktail/SOX10 Muscle Specific Actin

For each of the mainline screening immunomarkers in the table above, once a positive categorical origin is confirmed (lymphoma, carcinoma, melanoma, or sarcoma), additional markers will be tested to determine organ-specific origin of the primary tumor2-4.

Secondary Screening Panels for Tumors of Unknown Origin

Lymphoma Carcinoma Melanoma Sarcoma
CD20 [L26] Cytokeratin LMW (8/18) HMB45 Smooth Muscle Actin
PAX-5 p63 + CK5 / Desmoglein 3 + p40 (M) Tyrosinase Desmin
CD3 [LN10] Cytokeratin 7 / Cytokeratin 19 MART-1 Cocktail Myogenin
CD15 Cocktail / CD30 (Ki-1) CDX2 (M) + CDH17 (RM) / Cytokeratin 20 Pan Melanoma Cocktail-2 CD31
ALK [5A4] TTF-1 + Napsin A (RM) Microphthalmia Transcription Factor (MiTF) CD34
Kappa (M) + Lambda (P) Estrogen Receptor (ER) [SP1]* / Progesterone Receptor (PR) [16]* N/A CD99
CD68 / CD163 c-erbB-2/HER2 N/A ERG
CD10 Prostein + PSA + NKX3.1 / ERG N/A N/A
Cyclin D1 Synaptophysin N/A N/A
CD4 + CD8 PAX8 (M) N/A N/A
Bcl-2 / Bcl-6 [LN22] SALL4 N/A N/A
CD7 CD56 / MASH1 N/A N/A
Terminal Deoxynucleotidyl Transferase Hepatocyte Specific Antigen / Arginase-1 N/A N/A
MUM-1 / CD138 Glypican-3 (GPC3) N/A N/A
N/A GATA-3 + Uroplakin II N/A N/A

*These products are Research Use Only, not FDA cleared for clinical diagnosis.

Clinical References:

1. Wise J. BMJ. 2016 Jun 12; 353:i3277.
2. Selves J, et al. Cancers. 2018; 10(4):108.
3. Kandukuri SR, et al. Arch Pathol Lab Med. 2017 Aug; 141(8):1014-1032.
4. Chu P, Wu E, Weiss LM. Mod Pathol. 2000 Sep;13(9):962-72.

Biocare Medical Excellence in Patient Care Award at NSH 2018

Utah Gastroenterology is the 2018 recipient of the Biocare Medical Excellence in Patient Care Award. The Utah Gastroenterology Histology Lab is a relatively new, small, private practice histology lab in Salt Lake City. The lab was nominated by the group of pathologists that provide diagnosis for the biopsies taken at Utah Gastroenterology. The pathologists were impressed by the extra efforts the lab takes to provide them with information above and beyond what is needed to make an accurate diagnosis. The histology lab manages a complex workflow, accepting specimens from different locations across the state. Despite this, they are meticulous with their processes, and quality control.

Pictured from Left to Right: NSH President – Diane Sterchi, Laurie Peck – Utah Gastroenterology, Jesse Del Campo – Biocare Medical.

Multiplex IHC – Tech Talk

Multiplex IHC

Biocare™ Medical is the proven leader in providing Multiplex antibody cocktails and detection platforms that enable simultaneous IHC staining with multiple antibodies on a single slide. Our Multiplex detection system simplifies procedures by conserving precious tissue, saving valuable technician time, reducing the number of reagents and steps in the protocol, and allowing for simultaneous evaluation of multiple targets on one slide.

Prostate cancer stained with CK HMW + p63 + AMACR (RM)

Antibody Cocktails

Early on, Biocare recognized the market need for evaluating morphologically distinct markers to aid in solving complex clinical problems and simplifying interpretation, all while conserving precious patient tissue.

The most glaring clinical need was determined to be the ability to differentiate between prostatic intraepithelial neoplasia (PIN) and carcinoma of the prostate. There also needed to be a way to clearly identify any microinvasion or micrometastasis into adjacent prostate tissue. Additionally, this would need to be performed in a clinical laboratory, with superior accuracy and specificity. All of this whilst conserving the limited tissue (prostate needle biopsies are thin filaments of tissue) and reducing the time to result.

With these constraints and needs in mind, Biocare developed our flagship Multiplex IHC cocktail: CK HMW + p63 + AMACR (RM) [formerly known as PIN-4]. Studies have shown that combinations of CK HMW [34βE12], p63, and/or AMACR may be useful in the evaluation of normal prostate glands, prostatic intraepithelial neoplasia (PIN) and prostatic adenocarcinoma1,2,3. In prostate, CK HMW [34βE12] has been shown to be a useful marker of basal cells of normal glands and PIN, a precursor lesion to prostatic adenocarcinoma; whereas invasive prostatic adenocarcinoma typically lacks a basal cell layer3-5. p63, a homolog of the tumor suppressor p53, has been detected in nuclei of the basal epithelium in normal prostate glands; however, it was not expressed in malignant tumors of the prostate6. α-Methylacyl coenzyme A racemase (AMACR), also known as P504S, has been shown to be a specific marker of prostatic adenocarcinoma7-10. Additionally, prostate glands involved in PIN have been found to express AMACR, whereas AMACR was nearly undetectable in benign glands10-11.

  • Conserve tissues, time and money = Cost-effective
    • A single Multiplex IHC can replace up to 5 single Ab stains; reducing labor and reagent costs by at least 50%
  • Accurate analysis and easy interpretation of staining results
  • Increase predictive value by combining highly sensitive & highly specific antibodies on one slide
  • More rapidly exclude or diagnose disease states
  • Eliminate multiple slides to evaluate antigen ratios

Multiplex IHC Products

We have also developed Multiplex IHC antibody cocktails for additional tissue applications: Breast, Bladder, Lung, Skin, Colon, Lymphoma, Tumors of Unknown Origin, and Cell Death vs. Proliferation.

Tissue Multiplex Product
Prostate CK HMW + p63 + AMACR (RM)
Breast CK5/14 + p63 + CK7/18
Bladder Uro-2
Lung Desmoglein 3 + p40 (M) + Napsin A (RM)
Skin Pan Melanoma + Ki-67
Colon CDX2 (M) + CDH17 (RM)
Lymphoma Kappa (M) + Lambda (P)
Tumor of Unknown Origin CDX2 + CK7
Death vs. Proliferation Ki-67 + Caspase 3

Clinical References:
1. Shah RB, et al. Am J Clin Pathol. 2004 Oct; 122(4):517-23. 2. Sung MT, et al. Hum Pathol. 2007 Feb; 38(2):332-41. 3. Bostwick DG, Qian J. Mod Pathol. 2004 Mar; 17(3):360-79. 4. Humphrey PA. J Clin Pathol. 2007 Jan; 60(1):35-42. 5. Shah RB, et al. Am J Surg Pathol. 2002 Sep; 26(9):1161-8. 6. Signoretti S, et al. 2000 Dec; 157(6):1769-75. 7. Xu J, et al. Cancer Res. 2000 Mar 15; 60(6):1677-82. 8. Rubin MA, et al. JAMA. 2002 Apr 3; 287(13):1662-70. 9. Luo J, et al. Cancer Res. 2002 Apr 15; 62(8):2220-6. 10. Zhou M, et al. Am J Surg Pathol. 2002 Jul; 26(7):926-31. 11. Wu CL, et al. Hum Pathol. 2004 Aug; 35(8):1008-13.

Fixation on Histology Awards Biocare Medical & Jason Ramos Ph.D. Top 5 NSH Presentations!


According to Fixation on Histology, Biocare Medical’s Dr. Jason Ramos, Ph.D. was awarded one of the top 5 best presentations at NSH 2018. For full details read below!

“NSH’s Symposium/Convention is the largest histology conference in the United States with over 100 workshops, an exhibit hall, and plenty of opportunity to network with fellow histologists. If you’re considering attending, especially if it’s your first time, it might seem overwhelming. Fixation on Histology is here to help!

We’ve got the list of top 5 most popular workshops, at this year’s Convention.

5. Molecular Biology Principles and Techniques for the Histotech (WS#56): Molecular pathology is often a less familiar entity for those accustomed to IHC and proteins. This seminar is intended to provide a comprehensive overview of basic molecular biology to offer a better understanding of the applications and diagnostic relevance of molecular pathology. A review of the scientific principles of molecular biology will be conducted. This will include understanding the transcription and translation processes. Different molecular techniques from PCR to ISH will be covered. The pros and cons of these techniques will be discussed, including which molecular techniques and methods are best suited for diagnostic, screening or treatment assessment purposes. The presentation will also look at the future for molecular pathology, and discuss routes for personalized medicine.”
Molecular Biology Principles and Techniques for the Histotech

Biocare Medical PDL1 (CAL10) – Stands Up to the Competition


Several publications have compared Biocare’s PD-L1 (CAL10) antibody for immunohistochemistry to FDA approved PD-L1 clones SP263 and 28-8 in both rare and prevalent cancer types. Positive and significant correlation of expression, intensity and concordance resulted in both studies. Below is a recent abstract published in the Journal of Clinical Oncology using these different PD-L1 clones in malignant Thymoma and Thymic Carcinoma.  Also included is a link to a study published in Human Pathology using the different PD-L1 clones in Breast Cancer.


Immunohistochemical staining for programmed cell-death ligand 1 (PD-L1) in malignant thymoma and thymic carcinoma. Alexei Shimanovsky, Richard Cartun, Mary Fiel-Gan, Daniza Mandich, Jonathan Earle, Andrew L. Salner… Abstract e20003

Background: Recent development of anti-PD-1/L1 antibodies has demonstrated activity in various neoplasms. Thymic malignancies (TMS) are rare and treatment in advanced disease is limited. To evaluate the potential impact of anti-PD-1/L1 therapy in TMS, we examined the expression of PD-L1 in previously resected thymoma (TM) and thymic carcinoma (TC). Methods: We examined resected specimens from patients at Hartford Hospital with TM and TC between 2000 and 2014. Expression of PD-L1 was evaluated on formalin-fixed paraffin-embedded tissue. Immunohistochemical testing was done using four different clones of PD-L1 antibodies on the Leica Bond Max automated platform. The four clones include: E1L3N (Cell Signaling Technology), 28-8 (Epitomics) and SP142 (Spring Bioscience), and CAL10 (BioCare). PD-L1 expression was evaluated based on the percentage of tumor cells positive and their intensity graded as negative, weak (1+), moderate (2+), and strong (+3). The scoring was performed by three pathologists and was blinded for clinicopathologic data and antibody clones. Results: We evaluated a total of 29 patients, including 26 patients with TM and 3 with TC. Among the 29 available specimens, 12 had completed PD-L1 expression assessment at the time of submission. PD-L1 expression is present in 75-100% of the evaluated patients. All had positive PD-L1 staining by SP142 and CAL10. Three patients showed strong intensity by CAL10, and one by SP142. E1L3N and 28-8 had positive PD-L1 expression in 9 and 8 patients respectively with weak/moderate intensity. SP142 and CAL10 demonstrated the strongest concordance (R2 = 0.91) but there was significant variation between antibodies (R2 = 0.31-0.91). No correlation was detected between tumor grade and PD-L1 expression. There were focal areas that lacked expression in all of the evaluated specimens. Conclusions: There is increased expression of PD-L1 in TMS. The level of PD-L1 expression varies between the four PD-L1 antibodies. Increased PD-L1 expression provides evidence for the use of PD-L1 inhibitors in TMS. The variable staining highlights the heterogeneity of TMS and challenges in developing predictive biomarker in this cancer.

Citations: Immunohistochemical staining for programmed cell-death ligand 1 (PD-L1) in malignant thymoma and thymic carcinoma. Alexei Shimanovsky, Richard Cartun, Mary Fiel-Gan, Daniza Mandich, Jonathan Earle, Andrew L. Salner, Katrina Collins, Gregory Alan Otterson, and Benjamin F. Chu. Journal of Clinical Oncology 2017 35:15_suppl, e20003-e20003