BioREE-B™ Lanthanoid Staining Kit for Live Cell SEM Imaging

Complete kit

A fundamentally new sample-preparation method is now available as a practical laboratory tool

Research into lanthanoid staining has led to the development of a fundamentally new method for preparing biological samples for SEM.

BioREE-B makes this scientific development available for practical use in the laboratory.

BioREE-B is a reagent kit for two-stage supravital staining of biological samples. It enables the surface, the immediate subsurface layer, and elements of internal cellular structure to be examined without prior fixation, dehydration, or conductive coating.

BioREE-B Instructions for Use

BioREE-B Safety Data Sheet (SDS)

Reduces the time and number ofpreparation steps

Eliminates the need for prior chemical fixation in compatible applications

Supports work with samples that are sensitive to conventional preparation

Read the current Instructions for Use and SDS before use. Follow laboratory procedures for handling chemical reagents and biological samples, use personal protective equipment, and comply with storage and disposal requirements.

Tested by ZEISS Research Microscopy Solutions

A ZEISS specialist examined a tongue imprint containing cells and bacteria on a non-conductive plastic substrate. The sample was prepared with the BioREE lanthanoid staining kit to assess bacterial visibility and ultrastructural preservation after transport.

Imaging conditions: GeminiSEM 300, VPBSD detector, variable pressure mode, 15 kV, 20 Pa. The findings relate to the sample examined.

The analysis of prestained cells was possible in the FESEM without additional conduction or coating of the sample.

Ann-Katrin Unger 
ZEISS Research Microscopy Solutions, report author

Good BSE contrast. Bacterial colonies and labelled spots inside the cells were visible in the images.

Preservation after storage. After several days, some single bacteria with well-preserved ultrastructure could still be observed.

Ease of handling. The report describes the staining procedure as apparently easy to handle.

Read the full ZEISS report →

Applications

BioREE bridges fundamental science and solutions for multiple industries

We have turned an innovation into a practical tool

BioREE-B adds visual morphological data to quantitative methods and supports research into cells, microorganisms, and their interactions with materials

Pharmaceuticals and biotechnology

Research on strains, cell models, and adherent and suspension cultures. Visual data help compare how cells and microorganisms respond to experimental conditions and complement standard control methods.

Microbiology and cytology

Observation of cells, colonies, biofilms, cell-to-cell contacts, and interactions between tissues and microbiota. The method is suitable for research with impression smears, thin tissue samples, and microbial cultures.

Medical devices and biomaterials

Visual assessment of cell and microorganism adhesion to coatings, scaffolds, catheters, drains, and other complex surfaces. SEM images help analyze the morphology of interactions between a biological sample and a material.

Ophthalmology

Examination of the corneal and conjunctival epithelium, including samples obtained by impression cytology. BioREE-B enables visualization of cells and microorganisms present in the sample and provides data for an indirect comparative assessment of epithelial functional state.

Food industry and agricultural biotechnology

Research on yeasts, bacteria, and biofilms in applied microbiology. Visual information complements quantitative measurements and helps reveal population heterogeneity or surface condition.

Compatibility

Broad applicability — from cell cultures to surgical specimens

Adherent cell cultures, including cultures on three-dimensional carriers

01

Microbial cultures, including biofilm-forming cultures

02

Suspension cell cultures after placement on a solid support

03

Thin tissue samples, biopsy specimens, and surgical specimens

04

Impression smears from mucosal surfaces

05

Suitable
samples

Equipment requirements: A scanning electron microscope with a low-vacuum or variable-pressure mode and a backscattered-electron (BSE) detector. Operating parameters are selected according to the microscope model and sample type.

Discuss applications →

Conditions and limitations

• The method provides its highest information yield when used with live, unfixed cells.
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• The method is not intended for previously fixed tissue blocks.
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• Phosphate-based substrates are not recommended; bone and teeth are also unsuitable for examination.
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• The sample support must withstand mildly acidic conditions.

Comparison

Reduce the number of preparation steps and obtain more structural information

Conventional protocols for preparing biological samples for SEM typically include fixation, dehydration, drying, and conductive coating.

These steps make the sample compatible with the imaging conditions, but they complicate preparation and can introduce artifacts.

BioREE-B provides an additional research mode for applications where minimal sample deformation and richer morphological information are important.

Conventional protocols for preparing biological samples for SEM typically include fixation, dehydration, drying, and conductive coating.

These steps make the sample compatible with the imaging conditions, but they complicate preparation and can introduce artifacts.

BioREE-B provides an additional research mode for applications where minimal sample deformation and richer morphological information are important.

Comparison with conventional preparation

Multi-step fixation and dehydration

Two-stage staining of a live, unfixed sample

Conductive coating before examination

Examination of a compatible sample without prior coating

Coating affects the chemical analysis of the surface

The absence of a coating preserves more of the sample’s original information for microanalysis

Primary focus: surface microrelief

Surface, immediate subsurface layer, and elements of internal structure

Comparison with conventional preparation

Conventional approach

Conventional approach / Preparation with

Preparation with

Capabilities

Scientific principle

Expand the capabilities of SEM for studying biological samples

Compared with conventional sample-preparation methods, BioREE-B extends the capabilities of SEM by supplementing surface images with information about structures in the immediate subsurface layer.

• High-resolution visualization of ultrastructure in the immediate subsurface layer.
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• Identification of cells on complex objects and substrates through enhanced contrast.

• Comparative information about the functional state of cells within a single sample.
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• More accurate chemical microanalysis without prior conductive coating.

Contrast differences are used for comparative and indirect interpretation.

The method is not a stand-alone quantitative assay of metabolism, a method for species-level identification of microorganisms, or a clinical diagnostic test.

Sequential treatment with BioREE-B enhances compositional contrast in biological structures and expands the range of elements that can be distinguished in SEM images.

Results

More distinguishable structures mean more data for comparative analysis of a single sample

Sequential treatment with BioREE-B enhances compositional contrast in biological structures and expands the range of elements that can be distinguished in SEM images.

Two-stage staining makes more structures available for analysis

A colony of Staphylococcus aureus on the surface of a polypropylene catheter

A print of the tongue mucosa on a plastic surface with an adhesive coating: large cells of the desquamating epithelium are surrounded by bacterial colonies

A colony of Corynebacterium diphtheriae (biovar mitis) on laboratory plastic

A Staphylococcus aureus colony on the surface of a polypropylene catheter

A Salmonella sp. colony cultivated in a medium with a high glucose content on the surface of carbon tape

A human neutrophil phagocytosing a colony of Staphylococcus aureus cells on a culture dish

More distinguishable structures mean more data for comparative analysis of a single sample

Explore gallery

Principle

Scientific principle of the technology

BioREE-B combines lanthanoid staining with an additional treatment using lead salts. A live, unfixed sample is treated sequentially with two formulations that selectively label different biological structures.

As a result, the surface, the immediate subsurface layer, and selected elements of internal structure become accessible for examination in SEM images.

Contrast differences can be used for an indirect comparative assessment of cellular functional state.

Staining also stabilizes the cells’ natural dehydration and helps reduce sample deformation during examination.

Staining mechanism

Scientific publications

Explore mechanism

Using the kit

A sequential workflow — from sample preparation to SEM imaging

The workflow includes preparation of a compatible sample, two sequential staining stages, rinsing, and SEM imaging. If required, the post-fixation step specified in the instructions is added.

Check the compatibility of the sample, support, and equipment

Compatibility

01

Prepare the sample according to the instructions

Preparation

02

Perform the two staining stages in sequence

Staining

03

Complete the preparation and perform post-fixation if required

Final Preparation

04

Proceed to SEM imaging in an appropriate operating mode

SEM Imaging

05

Protocol for use

Rinse the sample initially with isotonic sodium chloride solution to remove growth-medium components and phosphate-containing tissue ground-substance fluids adsorbed on the sample surface, thereby preventing unwanted binding of lanthanoid and lead contrast ions, which have an affinity for phosphate.
Place the sample in an aqueous isotonic neodymium chloride solution (or another Ln3+ solution) and incubate for 30 minutes. Note: For best results, staining should be performed under conditions as close as possible to the physiological conditions for the cell type.
Perform the first intermediate rinse in distilled water to remove excess first contrast agent.
Place the sample in an aqueous isotonic lead acetate solution and incubate for 15 minutes. Note: Samples attached to substrates should be immersed upside down, without allowing the sample surface to touch the bottom of the container.
Perform the second intermediate rinse in distilled water to remove excess second contrast agent.
Place the sample in the fixation solution if long-term storage is required and/or when working with pathogenic organisms.
Rinse the sample with distilled water for 1–10 seconds to remove residual fixative.

Protocol for use

Evidence

The technology is described inscientific publications, protected by a patent, and tested on SEM equipment

Scientific publications, a patent, and independent testing confirm the technological foundation of the method

Feedback

BioREE-B is intended for in-house preparation ofbiological samples in laboratories equipped with a compatible scanning electron microscope

Start using BioREE-B in your laboratory

Submit an inquiry to learn more about thetechnology, clarify equipment requirements, andreceive current BioREE-B supply terms

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Start your project with Logix experts

BioREE-B™ is a lanthanoid staining technology for the study of live, unfixed cells

A fundamentally new sample-preparation method is now available as a practical laboratory tool

Tested by ZEISS Research Microscopy Solutions

BioREE bridges fundamental science and solutions for multiple industries

Pharmaceuticals and biotechnology

Microbiology and cytology

Medical devices and biomaterials

Ophthalmology

Food industry and agricultural biotechnology

Broad applicability — from cell cultures to surgical specimens

Reduce the number of preparation steps and obtain more structural information

Expand the capabilities of SEM for studying biological samples

Two-stage staining makes more structures available for analysis

Scientific principle of the technology

Staining mechanism

A sequential workflow — from sample preparation to SEM imaging

Compatibility

Preparation

Staining

Final Preparation

SEM Imaging

Protocol for use

The technology is described inscientific publications, protected by a patent, and tested on SEM equipment

Fast visualization of ocular surface microbiota

Rapid whole-cell sample preparation for SEM

Reversible metabolic arrest with lanthanoids

Technical testing on the ZEISS GeminiSEM 300

Start using BioREE-B in your laboratory

Submit an inquiry to learn more about thetechnology, clarify equipment requirements, andreceive current BioREE-B supply terms

hi@lgxtech.com