FISH-IF Quantification

FISH-IF QUANTIFICATION

(Designed for RNAscope®)

 

Product Overview

Indica Labs FISH-IF quantification module measures any number of fluorescently-labeled DNA/RNA ISH probes and immunofluorescent (IF) protein biomarkers on a cell-by-cell basis. This allows the user to rapidly contextualize the corresponding protein and gene expression profile of every cell across the tissue. HALO® FISH-IF analysis is designed to work with single or dual IHC-ISH assays including RNAscope™ and supports the H-score protocol for RNAscope as recommended by the manufacturer, ACD, a Bio-techne brand.


Advance your RNAscope image analysis by downloading the latest Quantitative RNAscope Image Analysis Guide from Indica Labs and ACD, a Bio-techne brand and view the latest high-impact publications using HALO RNAscope Image analysis.


An example of FISH-IF quantification is shown in the slider above.  Here, DAPI and an IF-labeled membrane biomarker (cyan) are used to identify cells of interest and the RNA FISH probe (pink) is measured in these cells.  Cells that express the membrane biomaker are highlighted in cyan.  The FISH probe spots are highlighted in pink. IF stains can be identified and quantified in any compartment – nucleus, cytoplasm and/or membrane.

The module reports numerous outputs including, but not limited to: number of IF positive cells in total and by cell compartment, Average IF intensity and average IF intensity by cell compartment, percentage IF positive total and by cell compartment, number and percentage of probe positive cells, probe copies per cell and area per cell, cell score for each probe (0, +1, +2, +3 and +4), and an H-score for each probe.  Additionally, users can define cell phenotypes based on IF and probe positivity.

The FISH-IF quantification 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.

FISH-IF QUANTIFICATION

(Designed for RNAscope®)

Product Overview

Indica Labs FISH-IF quantification module measures any number of fluorescently-labeled DNA/RNA ISH probes and immunofluorescent (IF) protein biomarkers on a cell-by-cell basis. This allows the user to rapidly contextualize the corresponding protein and gene expression profile of every cell across the tissue. HALO® FISH-IF analysis is designed to work with single or dual IHC-ISH assays including RNAscope™ and supports the H-score protocol for RNAscope as recommended by the manufacturer, ACD, a Bio-techne brand.


Advance your RNAscope image analysis by downloading the latest Quantitative RNAscope Image Analysis Guide from Indica Labs and ACD, a Bio-techne brand and view the latest high-impact publications using HALO RNAscope Image analysis.


An example of FISH-IF quantification is shown in the slider above.  Here, DAPI and an IF-labeled membrane biomarker (cyan) are used to identify cells of interest and the RNA FISH probe (pink) is measured in these cells.  Cells that express the membrane biomaker are highlighted in cyan.  The FISH probe spots are highlighted in pink. IF stains can be identified and quantified in any compartment – nucleus, cytoplasm and/or membrane.

The module reports numerous outputs including, but not limited to: number of IF positive cells in total and by cell compartment, Average IF intensity and average IF intensity by cell compartment, percentage IF positive total and by cell compartment, number and percentage of probe positive cells, probe copies per cell and area per cell, cell score for each probe (0, +1, +2, +3 and +4), and an H-score for each probe.  Additionally, users can define cell phenotypes based on IF and probe positivity.

RNAscope Data Analysis Guide by ACD, a Bio-techne brand

PUBLICATIONS

Here are a few publications that cite the use of our ISH-IHC and FISH-IF modules. Your publication not on the list?  Drop us an email to let us know about it!

TitleAuthorsYearJournalApplicationHALO Modules
Transcriptomic analysis links diverse hypothalamic cell types to fibroblast growth factor 1-induced sustained diabetes remissionBentsen MA, Rausch DM, Mirzadeh Z, Muta K, Scarlett JM, Brown JM, Herranz-Perez V, Baquero AF, Thompson J, Alonge KM, Faber CL, Kaiyala KJ, Bennett C, Pyke C, Ratner C, Egerod KL, Holst B, Meek TH, Kutlu B, Zhang Y, Sparso T, Grove KL, Morton GJ, Kornum BR, Garcia-Verdugo J-M, Secher A, Jorgensen R, Schwartz MW, Pers TH2020Nature CommunicationsMetabolismISH-IHC/FISH-IF
Detection of Epstein–Barr Virus in Periodontitis: A Review of Methodological ApproachesTonoyan L, Chevalier M, Vincent-Bungnas S, Marsault R, Doglio A2021MircroorganismsReviewISH/FISH, ISH-IHC/FISH-IF
Detection of engineered T cells in FFPE tissue by multiplex in situ hybridization and immunohistochemistryWright JH, Huang L-Y, Weaver S, Archila LD, McAfee MS, Hirayama AV, Chapuis AG, Bleakley M, Rongvaux A, Turtle CJ, Chanthaphavong S, Campbell JS, Pierce RH2020Journal of Immunological MethodsImmunologyISH/FISH, ISH-IHC/FISH-IF
Epicardium-derived cells organize through tight junctions to replenish cardiac muscle in salamanders Eroglu E, Yen C, Witman N, Elewa A, Araus A, Wang H, Szattler T, Umeano C, Sohlmer J, Goedel A, Simon A, Chien K
2022Nature Cell Biology
Myology
ISH/FISH, ISH-IHC/FISH-IF
Satellite repeat RNA expression in epithelial ovarian cancer associates with a tumor immunosuppressive phenotypePorter R, Sun S, Flores M, Berzolla E, You E, Phillips I, KC N, Desai N, Tai E, Szabolcs A, Lang E, Pankaj A, Raabe M, Thapar V, Xu K, Nieman L, Rabe D, Kolin D, Stover E, Pepin D, Stott S, Deshpande V, Liu J, Solovyov A, Matulonis U, Greenbaum B, Ting D2022The Journal of Clinical InvestigationImmuno-oncologyISH/FISH, ISH-IHC/FISH-IF
Systemic Nos2 Depletion and Cox inhibition limits TNBC disease progression and alters lymphoid cell spatial orientation and densitySomasundaram V, Ridnour L, Cheng R, Walke A, Kedei N, Bhattacharyya D, Wink A, Edmondson E, Butcher D, Warner A, Dorsey T, Scheiblin D, Heinz W, Bryant R, Kinders R, Lipkowitz S, Wong S, Pore M, Hewitt S, McVicar D, Anderson S, Chang J, Glynn S, Ambs S, Lockett S, Wink D2022Redox BiologyOncologyClassifer, ISH/FISH, Spatial Analysis, Registration, ISH-IHC/FISH-IF
Annexin A2/TLR2/MYD88 pathway induces arginase 1 expression in tumor-associated neutrophilsZhang H, Zhu X, Friesen T, Kwak J, Pisarenko T, Mekvanich S, Velasco M, Randolph T, Kargl J, Houghton A2022Journal of Clinical InvestigationImmuno-oncologySpatial Analysis, ISH-IHC/FISH-IF
Spatiotemporal transcriptome analysis reveals critical roles for mechano-sensing genes at the border zone in remodeling after myocardial infarctionYamada S, Ko T, Hatsuse S, Nomura S, Zhang B, Dai Z, Inoue S, Kubota M, Sawami K, Yamada T, Sassa T, Katagiri M, Fujita K, Katoh M, Ito M, Harada M, Toko H, Takeda N, Morita H, Aburatani H, Komuro I2022Nature Cardiovascular ResearchMyologyISH-IHC/FISH-IF
Loss of functional System x-c uncouples aberrant postnatal neurogenesis from epileptogenesis in the hippocampus of Kcna1-KO miceAloi M, Thompson S, Quartapella N, Noebels J2022Cell ReportsNeuroscienceISH-IHC/FISH-IF
Multiparameter immunohistochemistry analysis of HIV DNA, RNA and immune checkpoints in lymph node tissueRichardson Z, Deleage C, Tutuka C, Walkiewica M, Del Rio-Estrada P, Pascoe R, Evans V, Reyesteran G, Gonzales M, Roberts-Thomson S, Gonzalez-Navarro M, Torres-Ruiz, F, Estes J, Lewin S, Cameron P2021Journal of Immunological MethodsInfectious DiseaseISH-IHC/FISH-IF
The potassium channel auxiliary subunit Kv_2 (Kcnab2) regulates Kv1 channels and dopamine neuron firingYee J, Rastani A, Soden M2022Journal of NeurophysiologyNeuroscienceISH-IHC/FISH-IF
Transcriptional vulnerabilities of striatal neurons in human and rodent models of Huntington’s diseaseMatsushima A, Pineda S, Crittenden J, Lee H, Galani K, Mantero J, Tombaugh G, Kellis M, Heiman M, Graybiel A2023Nature CommunicationsNeuroscienceArea Quantification, ISH-IHC/FISH-IFHALO

Click here to initiate your free proof-of-concept HALO® image analysis.