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Custom Spatial Transcriptomic Probe Sets

Custom oligonucleotide probe sets for spatial transcriptomics, multiplex RNA imaging, and spatial genomics research. Bio-Synthesis supports MERFISH, seqFISH, smFISH, custom barcoded probe libraries, fluorescent labeling, conjugation chemistry, and advanced RNA imaging workflows.

MERFISH seqFISH smFISH Custom Panels

What Are Spatial Transcriptomic Probe Sets?

Spatial transcriptomics combines gene expression analysis with spatial information, helping researchers determine where RNA molecules are located within tissues, organs and individual cells.

Unlike bulk RNA sequencing or conventional single-cell RNA sequencing, spatial transcriptomic methods preserve tissue architecture while measuring transcript abundance. This makes it possible to connect RNA expression with cellular organization, tissue microenvironments, developmental processes, immune landscapes and disease pathology.

Bio-Synthesis manufactures custom oligonucleotide probe sets for spatial transcriptomic and RNA imaging workflows, ranging from single-molecule detection to highly multiplexed spatial genomics applications. Probe sets may include direct fluorescent labels, combinatorial barcodes, sequential readout sequences, amplification domains, haptens, spacers or error-correcting encoding strategies depending on the method.

Technology Guidance

Compare MERFISH, seqFISH and smFISH probe strategies for your spatial RNA imaging goals.

Custom Probe Architecture

Support for target-binding regions, barcode domains, readout sequences and fluorescent labels.

Manufacturing Support

Custom synthesis, fluorescent labeling, conjugation chemistry, probe pools and QC documentation.

Map

Spatial Context

Preserve tissue architecture while measuring RNA expression and transcript localization.

Cell

Single-Cell Resolution

Analyze RNA expression at cellular or subcellular resolution depending on the platform.

Mx

Multiplex Detection

Support targeted panels through high-plex RNA imaging strategies.

Panel

Custom Panels

Design marker panels around tissue type, disease model, pathway or cell identity.

Applications of Spatial Transcriptomics

Spatial transcriptomic probe sets are used wherever RNA expression must be interpreted within tissue structure and cellular context.

ONC

Oncology

Tumor microenvironment characterization, biomarker discovery, immune infiltration studies and therapy-response mapping.

NEU

Neuroscience

Neuronal subtype mapping, glial marker panels, brain architecture studies and disease-model profiling.

IMM

Immunology

Immune-cell localization, cytokine profiling, inflammation mapping and tissue immune landscapes.

DEV

Development & Atlases

Lineage programs, organoids, tissue patterning, cell atlas projects and spatial gene-expression maps.

MERFISH, seqFISH or smFISH: Which Probe Strategy Fits?

Spatial transcriptomics is a broad category. MERFISH, seqFISH and smFISH are related RNA imaging approaches, but each uses a different probe architecture and experimental strategy.

MERFISH — best for highly multiplexed spatial RNA imaging with error-correcting barcode architectures.

large panels
hundreds-thousands
encoding + readout

Probe Strategy

Encoding probes bind target RNA and carry readout sequences decoded through sequential imaging rounds.

Strengths

Error-robust barcode design, high multiplex capacity and single-cell spatial expression mapping.

Best Fit

Cell atlas studies, complex tissues, tumor microenvironment mapping and large gene panels.

seqFISH — best for sequential RNA imaging using combinatorial hybridization cycles.

sequential panels
hundreds-thousands
readout cycles

Probe Strategy

Transcripts are identified across multiple hybridization and imaging rounds using combinatorial fluorescence signatures.

Strengths

Flexible panel design, high multiplex capability and spatial expression profiling.

Best Fit

Tissue architecture, spatial gene expression mapping and high-content RNA imaging.

smFISH — best for direct visualization of individual RNA molecules with lower panel complexity.

RNA localization
1 to dozens
direct probes

Probe Strategy

Multiple short fluorescent probes hybridize to one transcript, creating punctate single-molecule signal.

Strengths

High sensitivity, direct transcript detection, lower complexity and strong validation utility.

Best Fit

RNA localization, expression validation, small targeted panels and mechanistic studies.

Choosing the Right Spatial Transcriptomic Probe Technology

Feature smFISH seqFISH MERFISH
Multiplexing Low to moderate High Very high
Typical Target Count Single genes to dozens Hundreds to thousands Hundreds to thousands+
Single-Molecule Detection Excellent Good Good
Experimental Complexity Lower Moderate to high High
Sequential Imaging Usually no Yes Yes
Error-Correcting Barcodes No Optional/custom Core feature

Common Probe Architecture Elements

T

Target-Binding Regions

Hybridize directly to RNA targets and define transcript specificity.

Bar

Barcode Domains

Encode transcript identity through combinatorial or sequential readout strategies.

RO

Readout Sequences

Enable secondary fluorescent probes to bind during imaging cycles.

FL

Fluorophores

Generate optical signals using FAM, Cy dyes, Alexa Fluor dyes, ATTO dyes, NIR dyes and more.

Amp

Amplification Elements

Support signal amplification, library handling or workflow-specific sensitivity improvement.

EC

Error Correction

Improve decoding accuracy in highly multiplexed MERFISH-like workflows.

Design Considerations for Spatial Transcriptomic Probe Sets

Panel performance depends on biological target selection, probe architecture, barcode orthogonality, fluorophore strategy, sample compatibility and manufacturing feasibility.

Target Selection — choose genes that answer the biological question and distinguish cell states.

Marker Genes

Select cell-type markers, pathway genes, disease markers or validation targets.

Expression Level

Low-abundance transcripts may require careful probe density, amplification or platform choice.

Isoforms

Define shared exons, isoform-specific regions or gene-family specificity.

Panel Size — match panel complexity to the spatial question and imaging platform.

Small Panels

Useful for validation, RNA localization and focused pathway studies.

Medium Panels

Support cell-type and pathway profiling with manageable imaging complexity.

High-Plex Panels

MERFISH and seqFISH strategies can support hundreds to thousands of transcripts.

Barcode Design — maintain orthogonality, decoding accuracy and readout compatibility.

Readout Domains

Barcoded readout sequences must be orthogonal and compatible with imaging cycles.

Error Correction

MERFISH-style designs may use error-robust barcodes to improve decoding reliability.

Manufacturing Review

Barcode and adapter regions should be checked for synthesis feasibility and cross-hybridization risk.

Fluorophore Strategy — choose dyes and conjugation chemistry around the microscope and multiplex plan.

Wide Dye Selection

Bio-Synthesis supports a broad range of fluorescent dyes, NIR dyes, specialty imaging dyes and multiplex-compatible fluorophores.

Conjugation Chemistry

Amino, thiol, biotin, DIG, DNP, spacers and custom conjugation strategies can support specialized workflows.

Channel Planning

Fluorophore choice should match lasers, filters, detector sensitivity and sample autofluorescence.

Sample Compatibility — consider tissue processing, fixation and RNA accessibility.

FFPE

Probe design may need to account for RNA fragmentation and fixation effects.

Fresh Frozen

Often useful for preserving RNA quality and tissue morphology.

Cultured Cells / Organoids

Support targeted RNA imaging, validation panels and mechanistic studies.

Custom Probe Manufacturing, Dyes, Conjugation Chemistry & QC

Bio-Synthesis supports spatial transcriptomic probe programs beyond basic oligo synthesis, including fluorescent labels, barcodes, readout probes, haptens, spacers, custom conjugation chemistry, purification and documentation.

Wide Fluorescent Dye Range

FAM, HEX, TAMRA, ROX, Cy3, Cy5, Alexa Fluor, ATTO, NIR dyes and specialty imaging labels.

Conjugation Chemistry

Amino, thiol, biotin, DIG, DNP, spacers, internal modified bases and custom dye conjugation strategies.

Barcode Integration

Readout domains, adapter sequences, encoding probes, sequential imaging designs and custom panel architecture.

Probe Pool Support

Custom oligo pools, normalized sets, plate formatting, sequence files and project-specific delivery formats.

Need a specific dye or chemistry? If your preferred fluorophore, hapten, spacer or conjugation handle is not listed, review the full dye and labeling capabilities here: Fluorescent-Labeled Oligonucleotides.

Spatial Transcriptomic Probe Set Workflow

From target selection and panel design through manufacturing, labeling, hybridization and imaging analysis.

01
Gene Selection

Choose marker genes, pathways, cell types or transcript panels.

02
Panel Design

Define MERFISH, seqFISH, smFISH or custom architecture.

03
Probe Manufacturing

Synthesize probe pools, readouts, encoding probes or labeled sets.

04
Labeling Strategy

Incorporate fluorophores, haptens, spacers or conjugation handles.

05
Hybridization

Apply probes to tissue, cells, organoids or prepared samples.

06
Imaging & Analysis

Generate spatial expression maps and transcript localization data.

FAQ

What is spatial transcriptomics?
Spatial transcriptomics measures RNA expression while preserving the spatial location of transcripts within cells or tissues.
What is the difference between MERFISH and seqFISH?
 MERFISH uses error-robust barcodes decoded through readout cycles, while seqFISH uses sequential hybridization rounds to assign transcripts through combinatorial fluorescence patterns.
What is the difference between MERFISH and smFISH?
 smFISH usually detects one or a small number of RNA targets directly with fluorescent probes, while MERFISH is designed for highly multiplexed RNA imaging using barcode and readout architectures.
Can Bio-Synthesis manufacture custom barcoded probe libraries?
 Yes. Bio-Synthesis supports custom oligonucleotide probe pools, barcode domains, readout probes, encoding probes and custom sequence architectures.
Are fluorescently labeled probes available?
 Yes. Bio-Synthesis supports a wide range of fluorescent dyes including FAM, Cy dyes, Alexa Fluor dyes, ATTO dyes, NIR dyes and specialty imaging fluorophores.
Which technology should I choose?
 Use smFISH for focused single-transcript or small-panel studies, seqFISH for sequential high-plex spatial expression mapping, and MERFISH for large error-robust multiplex RNA imaging panels.

Information Helpful for a Spatial Transcriptomics Probe Quote

Technology
MERFISH, seqFISH, smFISH
Panel
Gene list, target count
Architecture
Encoding, readout, direct
Labels
Dyes, haptens, handles
Sample
FFPE, frozen, cells
QC
HPLC, MS, CoA, docs

Need help selecting or manufacturing spatial transcriptomic probe sets?

Share your target gene list, technology preference, sample type, imaging platform, fluorophore requirements, barcode/readout design, conjugation chemistry, scale, purification and documentation needs.

Panel Strategy

MERFISH, seqFISH, smFISH, gene panels, barcode architecture and readout design.

MERFISH seqFISH smFISH

Labeling Options

Fluorescent dyes, haptens, spacers, amino/thiol handles and custom conjugation.

Dyes Barcodes QC

Quality Systems & Manufacturing Support

Spatial transcriptomic probe sets require controlled synthesis, sequence handling, labeling strategy, purification, documentation and project-specific delivery formats.

ISO-Supported Oligonucleotide Manufacturing Platform

Bio-Synthesis supports custom probe pool manufacturing, fluorescent labeling, barcode/readout oligos, custom conjugation chemistry, purification, analytical QC, documentation and packaging for spatial transcriptomics and RNA imaging workflows.

ISO 9001:2015 Quality management system
ISO 13485:2016 Medical-device quality framework
Analytical QC HPLC/UPLC, MS where compatible, OD260, CoA and traces
Custom Programs MERFISH, seqFISH, smFISH, RNA imaging and spatial biology probes

Spatial Transcriptomics Literature & Technical Background

  1. Ståhl PL, Salmén F, Vickovic S, et al. Visualization and analysis of gene expression in tissue sections by spatial transcriptomics. Science. 2016.
  2. Chen KH, Boettiger AN, Moffitt JR, Wang S, Zhuang X. Spatially resolved, highly multiplexed RNA profiling in single cells. Science. 2015.
  3. Lubeck E, Coskun AF, Zhiyentayev T, Ahmad M, Cai L. Single-cell in situ RNA profiling by sequential hybridization. Nature Methods. 2014.
  4. Eng CL, Lawson M, Zhu Q, et al. Transcriptome-scale super-resolved imaging in tissues by RNA seqFISH+. Nature. 2019.
  5. Raj A, van den Bogaard P, Rifkin SA, van Oudenaarden A, Tyagi S. Imaging individual mRNA molecules using multiple singly labeled probes. Nature Methods. 2008.
  6. Moses L, Pachter L. Museum of spatial transcriptomics. Nature Methods. 2022.

Technical note: Final probe design should be evaluated within the selected platform, target gene expression, sample preparation, imaging hardware, fluorophore channels, barcode architecture and data-analysis workflow.

Why Choose Bio-Synthesis

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