Multiplex FISH
MULTIPLEX FISH
(Designed for Fluorescent RNAscope®)
Product Overview
Indica Labs’ Multiplex FISH module quantifies an unlimited number of DNA or RNA FISH probes on a cell-by-cell basis. This analysis module can be used with most multiplex RNA FISH assays including RNAscope® and supports the H-score protocol for RNAscope® recommended by the manufacturer, ACD Bio.
Watch a webinar from two of our customers discussing how they use and analyze RNAscope assays in their research and download the full ACD Bio™ RNAscope Data Analysis Guide by clicking the links here.
In the example above the module measures a two-plex RNAscope assay on pelleted cancer cells. Single cells are identified using the nuclear dye DAPI. The multiplexed red and green probes are measured within the cell boundaries (spots per cell and probe area per cell). The module reports the average number of each probe per cell or across the entire region of analysis. In the analysis markup image, cells positive for the red probe alone are highlighted in different shades of red, cells positive for the green probe alone in different shades of green and cells positive for both probes are highlighted in shades of yellow. The color shading is determined by the number of probes and the corresponding RNAscope H-score, dark color = lower expression and light color = higher expression.
The Multiplex FISH module integrates seamlessly into the HALO® platform which is compatible with a number of third party systems and file formats.
Contact info@indicalab.com for product demonstration and pricing information or upload some images for a free trial.
MULTIPLEX FISH
(Designed for RNAscope®)
Indica Labs’ Multiplex FISH module quantifies an unlimited number of DNA or RNA FISH probes on a cell-by-cell basis. This analysis module can be used with most multiplex RNA FISH assays including RNAscope® and supports the H-score protocol for RNAscope® recommended by the manufacturer, Advanced Cell Diagnostics.
Watch a webinar from two of our customers discussing how they use and analyze RNAscope assays in their research and download the full ACD Bio™ RNAscope Data Analysis Guide by clicking the links here.
In the example above the module measures a two-plex RNAscope assay on pelleted cancer cells. Single cells are identified using the nuclear dye DAPI. The multiplexed red and green probes are measured within the cell boundaries (spots per cell and probe area per cell). The module reports the average number of each probe per cell or across the entire region of analysis. In the analysis markup image, cells positive for the red probe alone are highlighted in different shades of red, cells positive for the green probe alone in different shades of green and cells positive for both probes are highlighted in shades of yellow. The color shading is determined by the number of probes and the corresponding RNAscope H-score, dark color = lower expression and light color = higher expression.
HALO is compatible with a broad spectrum of image and digital slide formats. Yours not on the list? Email us your requirements.
Non-proprietary (JPG, TIF)
Nikon (ND2)
3D Histech (MRXS)
Perkin Elmer (QPTIFF, component TIFF)
Olympus (VSI)
Hamamatsu (NDPI, NDPIS)
Aperio (SVS, AFI)
Zeiss (CZI)
Leica (SCN, LIF)
Philips (Requires Philips IMS)
Quantitative RNAscope®️ Image Analysis Guide.
PUBLICATIONS
Here are a few publications that cite use of our ISH and FISH modules. Your publication not on the list? Drop us an email to let us know about it!
| Title | Authors | Year | Journal | Application | HALO Modules |
|---|---|---|---|---|---|
| Glucose-Dependent Insulinotropic Polypeptide Receptor-Expressing Cells in the Hypothalamus Regulate Food Intake | Adriaenssens AE, Biggs EK, Darwish T, Tadross J, Sukthankar T, Girish M, Polex-Wolf J, Lam BY, Zvetkova I, Pan W, Chiarugi D, Yeo GSH, Blouet C, Gribble FM, Reimann F | 2019 | Cell Metabolism | Metabolism | ISH/FISH |
| Fully automated RNAscope in situ hybridization assays for formalin‐fixed paraffin‐embedded cells and tissues | Anderson CM, Zhang B, Miller M, Butko E, Wu X, Laver T, Kernag C, Kim J, Luo Y, Lamparski H | 2016 | Journal of Cellular Biochemistry | Oncology, Gastroenterology, Infectious Disease | ISH/FISH |
| TGFβ inhibition restores a regenerative response in acute liver injury by suppressing paracrine senescence | Bird TG, Müller M, Boulter L, Vincent DF, Ridgway RA, Elena Lopez-Guadamillas E, Lu W-Y, Jamieson T, Govaere O, Campbell AD, Ferreira-Gonzalez S, Cole AM, Hay T, Simpson KJ, Clark W, Hedley A, Clarke M, Gentaz P, Nixon C, Bryce S, Kiourtis C, Sprangers J, Nibbs RJB, Van Rooijen N, Bartholin L, McGreal SR, Apte U, Barry ST, Iredale JP, Clarke AR, Serrano M, Roskams TA, Sansom OJ, Forbes SJ | 2018 | Science Translational Medicine | Metabolism | ISH/FISH |
| Food Perception Primes Hepatic ER Homeostasis via Melanocortin-Dependent Control of mTOR Activation | Brandt C, Nolte H, Henschke S, Ruud LE, Awazawa M, Morgan DA, Gabel P, Sprenger H-G, Hess ME, Günther S, Langer T, Rahmouni K, Fenselau H, Krüger M, Brüning JC | 2018 | Cell | Neuroscience, Metabolism | ISH/FISH |
| Detection and quantification of Parascaris P-glycoprotein drug transporter expression with a novel mRNA hybridization technique | Chelladurai JJ, Brewer MT | 2019 | Veterinary Parasitology | Infectious Disease | ISH/FISH |
| IL-7-dependent compositional changes within the γδ T cell pool in lymph nodes during ageing lead to an unbalanced anti-tumour response | Chen H-C, Eling N, Martinez-Jimenez CP, O'Brien LM, Carbonaro V, Marioni JC, Odom DT, de la Roche M | 2019 | EMBO Reports | Oncology, Immuno-oncology | ISH/FISH |
| Gene Signature of Proliferating Human Pancreatic α-Cells | Dominguez Gutierrez G, Xin Y, Okamoto H, Kim J, Lee A-H, Ni M, Adler C, Yancopoulos GD, Murphy AJ, Gromada J | 2018 | Endocrionology | Metabolism | ISH/FISH |
| Assessment of PD-L1 mRNA and protein expression in non-small cell lung cancer, head and neck squamous cell carcinoma and urothelial carcinoma tissue specimens using RNAScope and immunohistochemistry | Duncan DJ, Scott M, Scorer P, Barker C | 2019 | PLOS One | Oncology, Immuno-oncology | ISH/FISH |
| APNG as a prognostic marker in patients with glioblastoma | Fosmark S, Hellwege S, Dahlrot RH, Jensen KL, Derand H, Lohse J, Sørensen MD, Hansen S, Kristensen BW | 2017 | PLOS One | Oncology | ISH/FISH |
| Lipid Source and Peroxidation Status Alter Immune Cell Recruitment in Broiler Chicken Ileum | Fries-Craft KA, Meyer MM, Lindblom SC, Kerr BJ, Bobeck EA | 2021 | The Journal of Nutrition | Immunology | ISH/FISH |
| G-protein-independent coupling of MC4R to Kir7. 1 in hypothalamic neurons | Ghamari-Langroudi M, Digby GJ, Sebag JA, Millhauser GL, Palomino R, Matthews R, Gillyard T, Panaro BL, Tough IR, Cox HM, Denton JS, Cone RD | 2015 | Nature | Metabolism | ISH/FISH |
| A phase II study of the dual mTOR inhibitor MLN0128 in patients with metastatic castration resistant prostate cancer | Graham L, Banda K, Torres A, Carver BS, Chen Y, Pisano K, Shelkey G, Curley T, Scher HI, Lotan TL, Hsieh AC, Rathkopf DE | 2018 | Investigational New Drugs | Oncology | ISH/FISH |
| Expression of cholecystokinin by neurons in mouse spinal dorsal horn | Guitierrez-Mecinas M, Bell AM, Shepherd F, Polgar E, Watanabe M, Furuta T, Todd AJ | 2019 | The Journal of Comparitive Neurology | Neuroscience | ISH/FISH |
| Ultrasensitive automated RNA in situ hybridization for kappa and lambda light chain mRNA detects B-cell clonality in tissue biopsies with performance comparable or superior to flow cytometry | Guo L, Wang Z, Anderson CA, Doolittle E, Kernag S, Cotta CV, Ondrejka SL, Ma X-J, Cook JR | 2018 | Modern Pathology | Classifier, ISH/FISH | |
| A distinct repertoire of cancer-associated fibroblasts is enriched in cribriform prostate cancer | Hesterberg AB, Rios BL, Wolf EM, Tubbs C, Wong HY, Schaffer KR, Lotan TL, Giannico GA, Gordetsky JB, Hurley PJ | 2021 | Journal of Pathology:Clinical Research | Oncology | Classifier, ISH/FISH |
| Neurexin 3 transmembrane and soluble isoform expression and splicing haplotype are associated with neuron inflammasome and Alzheimer's disease | Hishimoto A, Pletnikova O, Lang DL, Troncoso JC, Egan JM, Liu Q-R | 2019 | Alzheimer's Research & Therapy | Neuroscience | ISH/FISH |
| Variability in Platelet-Derived Growth Factor Receptor Alpha Antibody Specificity May Impact Clinical Utility of Immunohistochemistry Assays | Holzer TR, Reising LO, Credille KM, Schade AE, Oakley | 2016 | Journal of Histochemistry & Cytochemistry | Oncology | Cytonuclear, ISH/FISH |
| Development of resistance to FAK inhibition in pancreatic cancer is linked to stromal depletion | Jiang H, Liu X, Knolhoff BL, Hegde S, Lee KB, Jiang H, Fields RC, Pachter JA, Lim K-H, DeNardo DG | 2019 | Gut | Oncology | Cytonuclear, ISH/FISH |
| Single-Cell Quantification of mRNA Expression in The Human Brain | Jolly S, Lang V, Koelzer VH, Frigerio CS, Magno L, Salinas PC, Whiting P, Palomer E | 2019 | Scientific Reports | Neuroscience | ISH/FISH |
| A first-in-human, phase 1, dose-escalation study of ABBV-176, an antibody-drug conjugate targeting the prolactin receptor, in patients with advanced solid tumors | Lemech C, Woodward N, Chan N, Mortimer J, Naumovski L, Nuthalapati S, Tong B, Jiang F, Ansell P, Ratajczak CK, Sachdev J | 2020 | Investigational New Drugs | Oncology | ISH/FISH |
| Mutant myocilin impacts sarcomere ultrastructure in mouse gastrocnemius muscle | Lynch JM, Dolman AJ, Guo C, Dolan K, Xiang C, Reda S, Li B, Prasanna G | 2018 | PLOS One | Gastroenterology, Myology | ISH/FISH |
| Breast and pancreatic cancer interrupt IRF8-dependent dendritic cell development to overcome immune surveillance | Meyer MA, Baer JM, Knolhoff BL, Nywening TM, Panni RZ, Su X, Weilbaecher KN, Hawkins WG, Ma C, Fields RC, Linehan DC, Challen GA, Faccio R, Aft RL, DeNardo DG | 2018 | Nature Communications | Oncology, Immuno-oncology | ISH/FISH |
| Gene expression of prostaglandin EP4 receptor in three canine carcinomas | Musser ML, Viall AK, Phillips RL, Hostetter JM, Johannes CM | 2020 | BMC Veterinary Research | Oncology | ISH/FISH |
| Cholecystokinin-1 receptor agonist induced pathological findings in the exocrine pancreas of non-human primates | Nyborg NCB, Kirk RK, de Boer AS, Andersen DW, Bugge A, Wulff BS, Thorup I, Clausen TR | 2020 | Toxicology & Applied Pharmacology | Metabolism | ISH/FISH |
| CD8+ T cells fail to limit SIV reactivation following ART withdrawal until after viral amplification | Okoye AA, Duell DD, Fukazawa Y, Varco-Merth B, Marenco A, Behrens H, Chaunzwa TM, Selseth AN, Gilbride RM, Shao J, Edlefsen PT, Geleziunas R, Pinkevych M, Davenport MP, Busman-Sahay K, Nekorchuk MD, Park H, Smedley JV, Axthelm MK, Estes JD, Hansen SG, Keele BF, Lifson JD, Picker LJ | 2021 | The Journal of Clinical Investigation | Immunology, Infectious Disease | ISH/FISH |
| Differential Cytokine Gene Expression in Granulomas from Lungs and Lymph Nodes of Cattle Experimentally Infected with Aerosolized Mycobacterium bovis | Palmer MV, Thacker TC, Waters WR | 2016 | PLOS One | Infectious Disease | ISH/FISH |
| The Fat Mass and Obesity-Associated Protein (FTO) Regulates Locomotor Responses to Novelty via D2R Medium Spiny Neurons | Ruud J, Alber J, Tokarska A, Engström Ruu L, Nolte H, Biglari N, Lippert R, Lautenschlager Ä, Cieślak PE, Szumiec Ł, Hess ME, Brönneke HS, Krüger M, Nissbrandt H, Korotkova T, Silberberg G, Rodriguez Parkitna J, Brüning J | 2019 | Cell Reports | Neuroscience, Metabolism | ISH/FISH |
| NPY mediates the rapid feeding and glucose metabolism regulatory functions of AgRP neurons | Ruud LE, Pereira MMA, de Solis AJ, Fenselau H, Bruning JC | 2020 | Nature Communications | Metabolism | ISH/FISH |
| Quantitative digital pathology reveals association of cell-specific PNPLA3 transcription with NAFLD disease activity | Sandhu B, Perez Matos MC, Tran S, Zhong A, Csizmadia E, Kim M, Herman MA, Nasser I, Lai M, Jiang ZG | 2019 | JHEP Reports | Fibrosis | ISH/FISH |
| Alpha-synuclein is a DNA binding protein that modulates DNA repair with implications for Lewy body disorders | Schaser AJ, Osterberg VR, Dent SE, Stackhouse TL, Wakeham CM, Boutros SW, Weston LJ, Owen N, Weissman TA, Luna E, Raber J, Luk KC, McCullough AK, Woltjer RL, Unni VK | 2019 | Scientific Reports | Neuroscience | ISH/FISH |
| Age-determined expression of priming protease TMPRSS2 and localization of SARS-CoV-2 infection in the lung epithelium | Schuler BA, Habermann AC, Plosa EJ, Taylor CJ, Jetter C, Kapp ME, Benjamin JT, Gullerman P, Nichols DS, Braunstein LZ, Hackett A, Koval M, Guttentag SH, Blackwell TS, Webber SA, Banovich NE, Kropski JA, Sucre JMS | 2020 | The Journal of Clincial Invesitgation | Infectious Disease | ISH/FISH |
| Small Molecule IL-36γ Antagonist as a Novel Therapeutic Approach for Plaque Psoriasis | Todorović V, Su Z, Putman CB, Kakavas SJ, Salte KM, McDonald HA, Wetter JB, Paulsboe SE, Sun Q, Gerstein CE, Medina L, Sielaff B, Sadhukhan R, Stockmann H, Richardson PL, Qiu W, Argiriadi MA, Henry RF, Herold JM, Shotwell JB, McGaraughty SP, Honore P, Gopalakrishnan SM, Sun CC, Scott VE | 2019 | Scientific Reports | Immunology, Dermatology | ISH/FISH |
| A randomized, open-label study of the efficacy and safety of AZD4547 monotherapy versus paclitaxel for the treatment of advanced gastric adenocarcinoma with FGFR2 polysomy or gene amplification | Van Cutsem E, Bang Y-J, Mansoor W, Petty RD, Chao Y, Cunningham D, Ferry DR, N. R. Smith NR, Frewer P, Ratnayake J, Stockman PK, Kilgour E, and Landers D | 2017 | Annals of Oncology | Oncology | Classifier, ISH/FISH |
| The human IL-15 superagonist N-803 promotes migration of virus-specific CD8+ T and NK cells to B cell follicles but does not reverse latency in ART-suppressed, SHIV-infected macaques | Webb GM, Molden J, Bushman-Sahay K, Abdulhaqq S, Wu HL, Weber WC, Bateman KB, Reed JS, Northrup M, Maier N,Tanaka S, Gao L, Davey B, Carpenter BL, Axthelm MK, Stanton JJ, Smedley J, Greene JM, Safrit JT, Estes JD, Skinner PJ, Sacha JB | 2020 | PLOS Pathogens | Infectious Disease | Cytonuclear, ISH/FISH |
| RNA Sequencing of Single Human Islet Cells Reveals Type 2 Diabetes Genes | Xin Y, Kim J, Okamoto H, Ni M, Wei Y, Adler C, Murphy AJ, Yancopoulos GD, Lin C, Gromada J | 2016 | Cell Metabolism | Metabolism | ISH/FISH |
| Expression of avian β-defensin mRNA in the chicken yolk sac | Zhang H, Wong EA | 2019 | Developmental & Comparative Immunology | Immunology | ISH/FISH |
| Activation of the renal GLP‐1R leads to expression of Ren1 in the renal vascular tree | Bjørnholm KD, Ougaard ME, Skovsted GF, Knudsen LB, Pyke C | 2021 | Endocrinology, Diabetes, & Metabolism | Metabolism | ISH/FISH |
| A Method to Pre-Screen Rat Mammary Gland Whole-Mounts Prior To RNAscope | Duderstadt EL, Sanders MA, Samuelson DJ | 2021 | Journal of Mammary Gland Biology and Neoplasia | ISH/FISH | |
| Age-associated expression of p21and p53 during human wound healing | Chia CW, Sherman-Baust CA, Larson SA, Pandey R, Withers R, Karikkineth AC, Zukley LM, Campisi J, Egan JM, Sen R, Ferrucci L | 2021 | Aging Cell | Dermatology | Area Quantification, ISH/FISH, Multiplex IHC |
| Immune-regulated IDO1-dependent tryptophan metabolism is source of one-carbon units for pancreatic cancer and stellate cells | Newman AC, Falcone M, Uribe AH, Zhang T, Athineos D, Pietzke M, Vazquez A, Blyth K, Maddocks ODK | 2021 | Molecular Cell | Immuno-oncology | Classifier, ISH/FISH |
| VGLUT2 is a determinant of dopamine neuron resilience in a rotenone model of dopamine neurodegeneration | Buck SA, De Miranda BR, Logan RW, Fish KN, Greenamyre JT, Freyberg Z | 2021 | The Journal of Neuroscience | Neuroscience | ISH/FISH |
| Mitigation of endemic GI-tract pathogen-mediated inflammation through development of multimodal treatment regimen and its impact on SIV acquisition in rhesus macaques | Bochart RM, Busman-Sahay K, Bondoc S, Morrow DW, Ortiz AM, Fennessey CM, Fischer MB, Shiel O, Swanson T, Shriver-Munsch CM, Crang HB, Armantrout KM, Barber-Axthelm AM, Langner C, Moats CR, Labriola CS, MacAllister R, Axthelm MK, Brenchley JM, Keele BF, Estes JD, Hansen SG, Smedley JV | 2021 | PLOS Pathogens | Infectious Disease | ISH/FISH, Multiplex IHC |
| Mucosal IFNγ production and potential role in protection in Escherichia coli O157:H7 vaccinated and challenged cattle | Schaut RG, Palmer MV, Boggiatto PM, Kudva IT, Loving CL, Sharma VK | 2021 | Scientific Reports | Immunology | ISH/FISH |
| Functionally distinct POMC-expressing neuron subpopulations in hypothalamus revealed by intersectional targeting | Biglari N, Gaziano I, Schumacher J, Radermacher J, Paeger L, Klemm P, Chen W, Corneliussen S, Wunderlich CM, Sue M, Vollmar S, Klöckener T, Sotelo-Hitschfeld T, Abbasloo A, Edenhofer F, Reimann F, Gribble FM, Fenselau H, Kloppenburg P, Wunderlich FT, Brüning JC | 2021 | Nature Neuroscience | Neuroscience | ISH/FISH |
Use the form below to upload up to three images for us to analyze using HALO. If the form does not display, please click here.