Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

Keynote webinar | Spotlight on medication adherence

LIVE: Thursday 27th June 2024, 18:00-19:30 (CEST)
Join our expert panel to discover why you need to understand the drivers of non-adherence in your patients, and how you can optimize medication adherence in your clinics to drastically improve patient outcomes.
ADVANCED SEARCH
Log in
Top
BMC Cancer
Published in: BMC Cancer 1/2008

Open Access 01-12-2008 | Research article

Comparison of mouse mammary gland imaging techniques and applications: Reflectance confocal microscopy, GFP Imaging, and ultrasound

Authors: Maddalena T Tilli, Angela R Parrish, Ion Cotarla, Laundette P Jones, Michael D Johnson, Priscilla A Furth

Published in: BMC Cancer | Issue 1/2008

Login to get access

Abstract

Background

Genetically engineered mouse models of mammary gland cancer enable the in vivo study of molecular mechanisms and signaling during development and cancer pathophysiology. However, traditional whole mount and histological imaging modalities are only applicable to non-viable tissue.

Methods

We evaluated three techniques that can be quickly applied to living tissue for imaging normal and cancerous mammary gland: reflectance confocal microscopy, green fluorescent protein imaging, and ultrasound imaging.

Results

In the current study, reflectance confocal imaging offered the highest resolution and was used to optically section mammary ductal structures in the whole mammary gland. Glands remained viable in mammary gland whole organ culture when 1% acetic acid was used as a contrast agent. Our application of using green fluorescent protein expressing transgenic mice in our study allowed for whole mammary gland ductal structures imaging and enabled straightforward serial imaging of mammary gland ducts in whole organ culture to visualize the growth and differentiation process. Ultrasound imaging showed the lowest resolution. However, ultrasound was able to detect mammary preneoplastic lesions 0.2 mm in size and was used to follow cancer growth with serial imaging in living mice.

Conclusion

In conclusion, each technique enabled serial imaging of living mammary tissue and visualization of growth and development, quickly and with minimal tissue preparation. The use of the higher resolution reflectance confocal and green fluorescent protein imaging techniques and lower resolution ultrasound were complementary.
Appendix
Available only for authorised users
Literature
1.
Chodosh LA, Cardiff RD: In vivo imaging of the mammary gland: the shape of things to come. J Mammary Gland Biol Neoplasia. 2006, 11: 101-102. 10.1007/s10911-006-9016-2.CrossRefPubMed
2.
Lyons SK: Advances in imaging mouse tumour models in vivo. J Pathol. 2005, 205: 194-205. 10.1002/path.1697.CrossRefPubMed
3.
Sidani M, Wyckoff J, Xue C, Segall JE, Condeelis J: Probing the microenvironment of mammary tumors using multiphoton microscopy. J Mammary Gland Biol Neoplasia. 2006, 11: 151-163. 10.1007/s10911-006-9021-5.CrossRefPubMed
4.
Sokolov K, Nieman LT, Myakov A, Gillenwater A: Polarized reflectance spectroscopy for pre-cancer detection. Technol Cancer Res Treat. 2004, 3: 1-14.CrossRefPubMed
5.
Farina KL, Wyckoff JB, Rivera J, Lee H, Segall JE, Condeelis JS, Jones JG: Cell motility of tumor cells visualized in living intact primary tumors using green fluorescent protein. Cancer Res. 1998, 58: 2528-2532.PubMed
6.
Massoud TF, Gambhir SS: Molecular imaging in living subjects: seeing fundamental biological processes in a new light. Genes Dev. 2003, 17: 545-580. 10.1101/gad.1047403.CrossRefPubMed
7.
Kavanaugh C, Green JE: The use of genetically altered mice for breast cancer prevention studies. J Nutr. 2003, 133: 2404S-2409S.PubMed
8.
Abbey CK, Borowsky AD, Gregg JP, Cardiff RD, Cherry SR: Preclinical imaging of mammary intraepithelial neoplasia with positron emission tomography. J Mammary Gland Biol Neoplasia. 2006, 11: 137-149. 10.1007/s10911-006-9020-6.CrossRefPubMed
9.
Wendel HG, de Stanchina E, Fridman JS, Malina A, Ray S, Kogan S, Cordon-Cardo C, Pelletier J, Lowe SW: Survival signalling by Akt and eIF4E in oncogenesis and cancer therapy. Nature. 2004, 428: 332-337. 10.1038/nature02369.CrossRefPubMed
10.
Rajadhyaksha M, Anderson RR, Webb RH: Video-Rate Confocal Scanning Laser Microscope for Imaging Human Tissues In Vivo. Applied Optics. 1999, 38: 2105-2115.CrossRefPubMed
11.
Lacy AA, Collier T, Price JE, Dharmawardhane S, Richards-Kortum R: Near real-time in vivo confocal imaging of mouse mammary tumors. Front Biosci. 2002, 7: f1-f7. 10.2741/lacy.CrossRefPubMed
12.
Demos SG, Vogel AJ, Gandjbakhche AH: Advances in optical spectroscopy and imaging of breast lesions. J Mammary Gland Biol Neoplasia. 2006, 11: 165-181. 10.1007/s10911-006-9022-4.CrossRefPubMed
13.
Gonzalez S, Swindells K, Rajadhyaksha M, Torres A: Changing paradigms in dermatology: confocal microscopy in clinical and surgical dermatology. Clin Dermatol. 2003, 21: 359-369. 10.1016/j.clindermatol.2003.08.007.CrossRefPubMed
14.
Tilli MT, Cabrera MC, Parrish AR, Torre KM, Sidawy MK, Gallagher AL, Makariou E, Polin SA, Liu MC, Furth PA: Real-Time Imaging and Characterization of Human Breast Tissue by Reflectance Confocal Microscopy. J Biomed Opt.
15.
Parrish A, Halama E, Tilli MT, Freedman M, Furth PA: Reflectance confocal microscopy for characterization of mammary ductal structures and development of neoplasia in genetically engineered mouse models of breast cancer. J Biomed Opt. 2005, 10: 051602-10.1117/1.2065827.CrossRefPubMed
16.
Chen CS, Elias M, Busam K, Rajadhyaksha M, Marghoob AA: Multimodal in vivo optical imaging, including confocal microscopy, facilitates presurgical margin mapping for clinically complex lentigo maligna melanoma. Br J Dermatol. 2005, 153: 1031-1036. 10.1111/j.1365-2133.2005.06831.x.CrossRefPubMed
17.
Collier T, Lacy A, Richards-Kortum R, Malpica A, Follen M: Near real-time confocal microscopy of amelanotic tissue: detection of dysplasia in ex vivo cervical tissue. Acad Radiol. 2002, 9: 504-512. 10.1016/S1076-6332(03)80326-4.CrossRefPubMed
18.
Nori S, Rius-Diaz F, Cuevas J, Goldgeier M, Jaen P, Torres A, Gonzalez S: Sensitivity and specificity of reflectance-mode confocal microscopy for in vivo diagnosis of basal cell carcinoma: a multicenter study. J Am Acad Dermatol. 2004, 51: 923-930. 10.1016/j.jaad.2004.06.028.CrossRefPubMed
19.
Hoffman RM: The multiple uses of fluorescent proteins to visualize cancer in vivo. Nat Rev Cancer. 2005, 5: 796-806. 10.1038/nrc1717.CrossRefPubMed
20.
Hadjantonakis AK, Gertsenstein M, Ikawa M, Okabe M, Nagy A: Generating green fluorescent mice by germline transmission of green fluorescent ES cells. Mech Dev. 1998, 76: 79-90. 10.1016/S0925-4773(98)00093-8.CrossRefPubMed
21.
Ballou B, Ernst LA, Waggoner AS: Fluorescence imaging of tumors in vivo. Curr Med Chem. 2005, 12: 795-805. 10.2174/0929867053507324.CrossRefPubMed
22.
Spergel DJ, Kruth U, Shimshek DR, Sprengel R, Seeburg PH: Using reporter genes to label selected neuronal populations in transgenic mice for gene promoter, anatomical, and physiological studies. Prog Neurobiol. 2001, 63: 673-686. 10.1016/S0301-0082(00)00038-1.CrossRefPubMed
23.
Yang M, Baranov E, Jiang P, Sun FX, Li XM, Li L, Hasegawa S, Bouvet M, Al Tuwaijri M, Chishima T, Shimada H, Moossa AR, Penman S, Hoffman RM: Whole-body optical imaging of green fluorescent protein-expressing tumors and metastases. Proc Natl Acad Sci USA. 2000, 97: 1206-1211. 10.1073/pnas.97.3.1206.CrossRefPubMedPubMedCentral
24.
Yang M, Baranov E, Li XM, Wang JW, Jiang P, Li L, Moossa AR, Penman S, Hoffman RM: Whole-body and intravital optical imaging of angiogenesis in orthotopically implanted tumors. Proc Natl Acad Sci USA. 2001, 98: 2616-2621. 10.1073/pnas.051626698.CrossRefPubMedPubMedCentral
25.
Manni A, Washington S, Craig L, Cloud M, Griffith JW, Verderame MF, Texter LJ, Mauger D, Demers LM, Harms JF, Welch DR: Effects of alpha-difluoromethylornithine on local recurrence and pulmonary metastasis from MDA-MB-435 breast cancer xenografts in nude mice. Clin Exp Metastasis. 2003, 20: 321-325. 10.1023/A:1024055522067.CrossRefPubMed
26.
Ray P, De A, Min JJ, Tsien RY, Gambhir SS: Imaging tri-fusion multimodality reporter gene expression in living subjects. Cancer Res. 2004, 64: 1323-1330. 10.1158/0008-5472.CAN-03-1816.CrossRefPubMedPubMedCentral
27.
Matulka LA, Triplett AA, Wagner KU: Parity-induced mammary epithelial cells are multipotent and express cell surface markers associated with stem cells. Dev Biol. 2007, 303: 29-44. 10.1016/j.ydbio.2006.12.017.CrossRefPubMed
28.
Ahmed F, Wyckoff J, Lin EY, Wang W, Wang Y, Hennighausen L, Miyazaki J, Jones J, Pollard JW, Condeelis JS, Segall JE: GFP expression in the mammary gland for imaging of mammary tumor cells in transgenic mice. Cancer Res. 2002, 62: 7166-7169.PubMed
29.
Yang M, Li L, Jiang P, Moossa AR, Penman S, Hoffman RM: Dual-color fluorescence imaging distinguishes tumor cells from induced host angiogenic vessels and stromal cells. Proc Natl Acad Sci USA. 2003, 100: 14259-14262. 10.1073/pnas.2436101100.CrossRefPubMedPubMedCentral
30.
Yang M, Jiang P, Hoffman RM: Whole-body subcellular multicolor imaging of tumor-host interaction and drug response in real time. Cancer Res. 2007, 67: 5195-5200. 10.1158/0008-5472.CAN-06-4590.CrossRefPubMed
31.
Coatney RW: Ultrasound imaging: principles and applications in rodent research. ILAR J. 2001, 42: 233-247.CrossRefPubMed
32.
Weissleder R: Scaling down imaging: molecular mapping of cancer in mice. Nat Rev Cancer. 2002, 2: 11-18. 10.1038/nrc701.CrossRefPubMed
33.
Boone JM, Kwan AL, Yang K, Burkett GW, Lindfors KK, Nelson TR: Computed tomography for imaging the breast. J Mammary Gland Biol Neoplasia. 2006, 11: 103-111. 10.1007/s10911-006-9017-1.CrossRefPubMed
34.
Rooks V, Beecken WD, Iordanescu I, Taylor GA: Sonographic evaluation of orthotopic bladder tumors in mice treated with TNP-470, an angiogenic inhibitor. Acad Radiol. 2001, 8: 121-127. 10.1016/S1076-6332(01)90038-8.CrossRefPubMed
35.
Oelze ML, O'Brien WD, Blue JP, Zachary JF: Differentiation and characterization of rat mammary fibroadenomas and 4T1 mouse carcinomas using quantitative ultrasound imaging. IEEE Trans Med Imaging. 2004, 23: 764-771. 10.1109/TMI.2004.826953.CrossRefPubMed
36.
Graham KC, Wirtzfeld LA, MacKenzie LT, Postenka CO, Groom AC, MacDonald IC, Fenster A, Lacefield JC, Chambers AF: Three-dimensional high-frequency ultrasound imaging for longitudinal evaluation of liver metastases in preclinical models. Cancer Res. 2005, 65: 5231-5237. 10.1158/0008-5472.CAN-05-0440.CrossRefPubMed
37.
Ferrara KW, Merritt CR, Burns PN, Foster FS, Mattrey RF, Wickline SA: Evaluation of tumor angiogenesis with US: imaging, Doppler, and contrast agents. Acad Radiol. 2000, 7: 824-839. 10.1016/S1076-6332(00)80631-5.CrossRefPubMed
38.
Sehgal CM, Weinstein SP, Arger PH, Conant EF: A review of breast ultrasound. J Mammary Gland Biol Neoplasia. 2006, 11: 113-123. 10.1007/s10911-006-9018-0.CrossRefPubMed
39.
Le Pivert P, Haddad RS, Aller A, Titus K, Doulat J, Renard M, Morrison DR: Ultrasound guided combined cryoablation and microencapsulated 5-Fluorouracil inhibits growth of human prostate tumors in xenogenic mouse model assessed by luminescence imaging. Technol Cancer Res Treat. 2004, 3: 135-142.CrossRefPubMed
40.
Tilli MT, Frech MS, Steed ME, Hruska KS, Johnson MD, Flaws JA, Furth PA: Introduction of estrogen receptor-alpha into the tTA/TAg conditional mouse model precipitates the development of estrogen-responsive mammary adenocarcinoma. Am J Pathol. 2003, 163: 1713-1719.CrossRefPubMedPubMedCentral
41.
Tzeng YJ, Guhl E, Graessmann M, Graessmann A: Breast cancer formation in transgenic animals induced by the whey acidic protein SV40 T antigen (WAP-SV-T) hybrid gene. Oncogene. 1993, 8: 1965-1971.PubMed
42.
Li M, Hu J, Heermeier K, Hennighausen L, Furth PA: Apoptosis and remodeling of mammary gland tissue during involution proceeds through p53-independent pathways. Cell Growth Differ. 1996, 7: 13-20.PubMed
43.
Fraschini A, Pellicciari C, Biggiogera M, Manfredi Romanini MG: The effect of different fixatives on chromatin: cytochemical and ultrastructural approaches. Histochem J. 1981, 13: 763-769. 10.1007/BF01003288.CrossRefPubMed
44.
Smithpeter CL, Dunn AK, Drezek RA, Collier TG, Richards-Kortum RR: Near real time confocal microscopy of cultured amelanotic cells: sources of signal, contrast agents and limits of contrast. Journal of Biomedical Optics. 1998, 3: 429-436. 10.1117/1.429853.CrossRefPubMed
45.
Rajadhyaksha M, Zavislan JM: Confocal laser microscope images tissue in vivo. Laser Focus World. 1997, 33: 5-8.
46.
Ginsburg E, Vonderhaar BK: Whole organ culture of the mouse mammary gland. Methods in Mammary Gland Biology and Breast Cancer Research. Edited by: Ip MM, Asch BB. 2000, New York, NY: Kluwer Academic/Plenum Publishers, 147-154.CrossRef
47.
Gouon-Evans V, Pollard JW: Unexpected deposition of brown fat in mammary gland during postnatal development. Mol Endocrinol. 2002, 16: 2618-2627. 10.1210/me.2001-0337.CrossRefPubMed
48.
Li X, Wang J, An Z, Yang M, Baranov E, Jiang P, Sun F, Moossa AR, Hoffman RM: Optically imageable metastatic model of human breast cancer. Clin Exp Metastasis. 2002, 19: 347-350. 10.1023/A:1015510207431.CrossRefPubMed
49.
Campo-Ruiz V, Lauwers GY, Anderson RR, Delgado-Baeza E, Gonzalez S: In vivo and ex vivo virtual biopsy of the liver with near-infrared, reflectance confocal microscopy. Mod Pathol. 2005, 18: 290-300. 10.1038/modpathol.3800303.CrossRefPubMed
50.
Clark AL, Gillenwater AM, Collier TG, Alizadeh-Naderi R, El Naggar AK, Richards-Kortum RR: Confocal microscopy for real-time detection of oral cavity neoplasia. Clin Cancer Res. 2003, 9: 4714-4721.PubMed
51.
Drezek RA, Richards-Kortum R, Brewer MA, Feld MS, Pitris C, Ferenczy A, Faupel ML, Follen M: Optical imaging of the cervix. Cancer. 2003, 98: 2015-2027. 10.1002/cncr.11678.CrossRefPubMed
52.
Ciarloni L, Mallepell S, Brisken C: Amphiregulin is an essential mediator of estrogen receptor alpha function in mammary gland development. Proc Natl Acad Sci USA. 2007, 104: 5455-5460. 10.1073/pnas.0611647104.CrossRefPubMedPubMedCentral
53.
Hoffman RM: Imaging tumor angiogenesis with fluorescent proteins. APMIS. 2004, 112: 441-449. 10.1111/j.1600-0463.2004.apm11207-0806.x.PubMed
54.
Li Y, Gonzalez S, Terwey TH, Wolchok J, Li Y, Aranda I, Toledo-Crow R, Halpern AC: Dual mode reflectance and fluorescence confocal laser scanning microscopy for in vivo imaging melanoma progression in murine skin. J Invest Dermatol. 2005, 125: 798-804. 10.1111/j.0022-202X.2005.23786.x.CrossRefPubMed
55.
Abramoff MD, Magelhaes PJ, Ram SJ: Image Processing with ImageJ. Biophotonoics International. 2004, 11: 36-42.
Metadata
Title
Comparison of mouse mammary gland imaging techniques and applications: Reflectance confocal microscopy, GFP Imaging, and ultrasound
Authors
Maddalena T Tilli
Angela R Parrish
Ion Cotarla
Laundette P Jones
Michael D Johnson
Priscilla A Furth
Publication date
01-12-2008
Publisher
BioMed Central
Published in
BMC Cancer / Issue 1/2008
Electronic ISSN: 1471-2407
DOI
https://doi.org/10.1186/1471-2407-8-21

Other articles of this Issue 1/2008

BMC Cancer 1/2008 Go to the issue

Research article

Leptin/HER2 crosstalk in breast cancer: in vitro study and preliminary in vivoanalysis

Research article

Detection of mismatch repair gene germline mutation carrier among Chinese population with colorectal cancer

Research article

Bioinformatic identification of FGF, p38-MAPK, and calcium signalling pathways associated with carcinoma in situin the urinary bladder

Technical advance

A novel circular invasion assay mimics in vivo invasive behavior of cancer cell lines and distinguishes single-cell motility in vitro

Research article

Pancreatobiliary versus intestinal histologic type of differentiation is an independent prognostic factor in resected periampullary adenocarcinoma

Research article

Nuclear hBD-1 accumulation in malignant salivary gland tumours
Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras
Prof. Fabrice Barlesi
Developed by: Springer Medicine
Image Credits