BIOREADER® 7000 -F-z-i micro

Versatile inverted Fluoro/VIS Reader, micro + macro zoom, high power LED for fluoro applications

BIOREADER® 7000 -F-z-i micro
Versatile inverted Fluoro/VIS Reader, micro + macro zoom, high power LED for fluoro applications

The Bioreader® 7000 -F-z-i micro provides an automatic zoom range for full-field-of-view of 35 - 100 mm Petridish, 6 - 384 well plates and max. 1 x 1 mm zoom factor on microscopic slides or hemocytometer.

Optical resolution for good shape recognition: Recommended from 10 µm(1) for single cells, 10 - 2000 µm for FOCI, 100 - 5000 µm for plaques or 50 - 2000 µm for cell clones.

(1) Depending on the selected front lens on the micro zoom (2x, 3x or 4x).

Model types          Fluoro  lightsources      Remission filters installed      VIS lightsources

Alpha                                           1                                             1                                        3

Beta                                             2                                             2                                        3

Gamma                                       3                                             3                                        3

Delta                                          6-8                                          6-8                                      3

 

Flexibility

The Bioreader® hardware may be upgraded on site. Brickstone principle.

Independence

Bioreader® operates independent form any assays or kit manucafurer, so you may choose among all assay/kit manufactures.

 

 

Antiviral vaccine development by use of the Bioreader®

This Bioreader ® supports testing and development of  antiviral vaccines.

Example#1: The automatic analyses of   Plaque reductions assay.

This assay applies diluted doses of a -possibly- antiviral drug into microwell plates. The Bioreader automatically observes its capability to suppress the formation of plaques(-holes-) in the bacterial lawn.

The ‘Plaque reductions neutralization test’ (PRNT) is a variation of this test. It is an attempt to find an antibody that neutralizes a specific virus. This is a very valuable tool in antiviral vaccine research.

Examplel#2: The automatically evaluation of the Yield reduction assay

In several steps of operation cell are infected in presence of variable doses of a possibly antiviral drug substance.

Thereafter the virus titer is determined in the liquid by use of the plaque or TCID50 assay with of  the Bioreader®.

The titer is calculated as the quotient of the positive cells/all cells, taking in account the virus dilution factor and the volume of the inoculum.

Features
  • Full automation of all mechanical functions incl. focus, zoom, aperture.
  • 8 times filter changer.
  • Upgrade option from model alpha to gamma and delta.
  • Dual telecentric illumination for better cytokine quantification.
  • Automated zoom lens for 6-48 well plates and/or Petri dishes.
  • Fixed focus lens for 96-384 well plates or Petri dishes.
  • Microscopic zoom system for microscopic cell counting.
  • Fluoro/UV LED excitation plus 3 types of white light LED illuminations.
  • Hardware is automatically controlled by software measure setup (GxP-locked).
  • No mercury or deuterium lamps required, thus 50T hourly lifetime.
  • Improved suppression of auto-fluorescence, cluster separation, well recognition and cytokine volume quantification.
  • Brand-new generation 7 makes it significant faster and more precise regarding positioning.
  • Single click export to PDF, Microsoft Office® applications.
  • Front loader, automatic door.
  • Dual tele-centric illumination.
  • Loud speaker and interactive training program.
  • Excel/Word/Power Point and LIMS export capabilities.
  • Suitable for research purpose.
  • High contrast/low noise cameras 5-41 MPixel available.
  • Optional full automation with plate 'feeding system'.
  • DQ/IQ/OQ/PQ documention possible.
  • 12 month warranty.
Software

biocreader_7000_fzi_micro_software.png

Eazyreader® software

  • Innovative EazyReader® software combines ‘easy of use’ and versatility and flexibility.
  • Scan, analyze and overlay live time.
  • Creates up to 7 images for each well simultaneously during the scan.
  • ‘Profiling’ app helps to create user independent measure protocols for Elispot beginners and references for experts.
  • Verified in collaborative studies.
  • Export options and reports with all scalable images and results even in one file.
  • Customer specific export and report templates.
  • Video clips and content specific help files.
  • Qualified installation and training with each Bioreader® model.
  • On-site or internet remote services and support.
  • ‘Classified’ measure protocols ‘history’ tracking and comparison tools.
  • User specific plates, Studies/projects, plate layouts/designs and measure protocols prevent from mix-up.
  • More accurate ‘cytokine quantification’ based on the patented ‘photometric’ dual illumination system.
  • Optional software features:
  • 21 CFR part 11 based software module available

Dimension

width
60 cm
height
80 cm
depth
45 cm

Weight

weight
40 kg
The BIOREADER® 7000 -F-z-i micro can also use with the follow applications:
Cell compartment visualization and cell transfection efficiency Icon

Cell compartment visualization and cell transfection efficiency

Cell compartment visualization and cell transfection efficiency

It is possible by use of the Bioreader® 7000-F-z with micro-resolution to determine the transfection rate. Differciation of compartments should be possible if compartments are transfected with specific proteins and labelled with individual fluorophores.

Elispot in 384 microfilter plates Icon

Elispot in 384 microfilter plates

Small volume, higher throughput.

Elispot - enzymatic single color Icon

Elispot - enzymatic single color

Elispot single encymatic

Blue, red, green or silver substrat Elispot.

Detection antibody, enzyme-conjugate and precipitating substrate.

"The enzyme-linked immune absorbent spot (ELISpot) is a type of assay that focuses on quantitatively measuring the frequency of cytokine secretion for a single cell."

[https://en.wikipedia.org/wiki/ELISpot]

Why to use Elispot

•The cytokine Elispot assay is designed to enumerate cytokine secreting cells in single cell suspensions of lymphoid tissue, CNS tissue, bone marrow or preparations of peripheral blood mononuclear cells (PBMC).

•The assay has the advantage of detecting only activated/memory T cells and the cytokine release can be detected at the single cell level, allowing direct determination of T cell frequencies.

•The assay has the advantage of detecting only activated/memory T cells and the cytokine release can be detected at the single cell level, allowing direct determination of T cell frequencies.

The ELISPOT assay is an effective tool to enumerate antigen-specific T cells in the circulation of immunized humans and animals at much lower frequencies than possible with other currently available methods

The ELISPOT assay has proven to be a sensitive and unique system to follow disease progression in human individuals or animals. Several studies have indicated that alterations in the frequency of cytokine pc in different compartments of the body adequately reflect changes in immune function

The ELISPOT assay may be used to determine effects of drugs, chemicals or other compounds on cytokine secretion in vitro, thereby providing data on their putative modulatory effects on immune function in vivo

Fluorospot 1-4 fluorophores Icon

Fluorospot 1-4 fluorophores

Multi Fluoro Color Reader,

Best for Elispot and FluoroSpot.

 

Applies ‘local neighborhood processing’ in order to compensate for uneven background on fluoro plates. This way the spot volume is quantified, independent of the local background.

 

Why use Fluorospot?

Enzymatic Elispot labeling
• allows to label 1-2 cytokines per cell simultanously.
• If you are interested to see ‘double secreting’ cells the stain of two cells in neighborhood may overlap. So, It is not clear if two cells are overlapping or one cells is really secreting two different cytokines ( see below left)

 

‘Fluorospot’ labeling
Allows to label  -a theoretical unlimited number different-  cytokines. Because excitation for each cytokine is applied -one after the other-, decisions for double secreting cells is clearly.
One Image is acquired for each cytokine!

Currently Fluorospot assays with 1-4 Fluorophores are on the market.

Fluorospot offer's significant advantages over colorimetric formats, particularly in the areas of multiplexing and automated spot detection.

Moreover, as spot development is not enzymatic, signal intensity is directly proportional to the amount of analyte within.

 

Infection assay Icon

Infection assay

Cell infection assay

Plaque reduction assay and Yield reduction assay are approved methods to determine antiviral substances

Verify virus titer by use of TCID50 (Median Tissue Culture Infectious Dose)

Example: Green fluorescence. Adherent cells (primary human fibroblasts) in 96-well-mikrotiterplate. Fluorophores: DAPI and AlexaFluor488.

Microscopic magnification for immunohistochemical and  fluorophores staining for plates from 6 to 384 wells and Microscope/Hemocytometer  slides.

>

GFP/DAPI-expressed virus infected Icon

GFP/DAPI-expressed virus infected

GFP/DAPI-expressed virus infected

"Using DNA recombinant technology, scientists combine the GFP gene to a another gene that produces a protein that they want to study, and then they insert the complex into a cell." [https://embryo.asu.edu/pages/green-fluorescent-protein]

RFP-expressed virus infected Icon

RFP-expressed virus infected

RFP-expressed virus infected

"Using DNA recombinant technology, scientists combine the RFP gene to a another gene that produces a protein that they want to study, and then they insert the complex into a cell." [https://embryo.asu.edu/pages/green-fluorescent-protein but with RFP instead of GFP]

non stained cells Icon

non stained cells

Non stained cells may be counted in a kind of phase contrast mode.

Alveolar epithelial cells (AEC) Icon

Alveolar epithelial cells (AEC)

Alveolar epithelial cells (AEC)

"Typically, type 1 alveolar cells comprise the major gas exchange surface of the alveolus and are integral to the maintenance of the permeability barrier function of the alveolar membrane. Type 2 pneumocytesare the progenitors of type 1 cells and are responsible for surfactant production and homeostasis." [https://www.sciencedirect.com/topics/medicine-and-dentistry/alveolar-type-i-cells]

The alveolar epitheliumis a major target in toxic exposures of the lung because of its structural delicacy and proximity to inhaled toxicants. Type II epithelial cellsare important in maintaining the integrity of alveolar epitheliumand normal lung function.

Elispot - enzymatic dual color Icon

Elispot - enzymatic dual color

Blue and red Elispot substrate (mixed color violet)

Dual cytokine secretion.

Cell viability Icon

Cell viability

Cell viability

6-CF appears as green fluorescence.

Live/dead Icon

Live/dead

Live/dead cells

Live cells are white, dead cells are black. Tryphaneblue stain.

Transgene assay Icon

Transgene assay

Transgene assay

"In its most precise usage, the term transgenedescribes a segment of DNA containing a gene sequence that has been isolated from one organism and is introduced into a different organism." [https://en.wikipedia.org/wiki/Transgene]

Cells transfected with an expression plasmid encoding eGFP.

Live single cells Icon

Live single cells

Live single cells

Bright field application Jurkatcells.

Live/apoptotic cells Icon

Live/apoptotic cells

Live/apoptotic cells

NUCblue/interference/NUCgreen. Mix of Jurkatcells untreated cell proliferate and TNF/CHX-treated that causes to apoptosis.

MCMV (Murine Cytomegalovirus) greenfluoro remission Icon

MCMV (Murine Cytomegalovirus) greenfluoro remission

MCMV (Murine Cytomegalovirus)

green fluoro remission

"Cytomegalovirus (CMV) (from the Greek cyto-, "cell," and megalo-, "large") is a genus of viruses in the order Herpesvirales." [https://en.wikipedia.org/wiki/Cytomegalovirus]

MCMV (Murine Cytomegalovirus) red fluoro remission. Icon

MCMV (Murine Cytomegalovirus) red fluoro remission.

MCMV (Murine Cytomegalovirus)

red fluoro remission

MCMV: mouse cytomegalovirus, unstained Icon

MCMV: mouse cytomegalovirus, unstained

MCMV: mouse cytomegalovirus, unstained

HCMV Human cytomegalovirus, unstained Icon

HCMV Human cytomegalovirus, unstained

HCMV human cytomegalovirus, unstained

"The glycoprotein gO (UL74) of human cytomegalovirus (HCMV) forms a complex with gH/gL. Virus mutants with a deletion of gO show a defect in secondary envelopment with the consequence that virus spread is restricted to a cell-associated pathway." [https://www.ncbi.nlm.nih.gov/pubmed/20181688]

MCMV mutants got generated that express with tdTomato, eGFP or mCherry Icon

MCMV mutants got generated that express with tdTomato, eGFP or mCherry

MCMV mutants got generated that express with tdTomato, eGFP or mCherry

MCMV mutants, tdTomato, Cherry, eGFP fibroplasts

NK92 cells natural killer cells (human cell line) expressing green fluorescent protein (gfp) Icon

NK92 cells natural killer cells (human cell line) expressing green fluorescent protein (gfp)

NK92 cells natural killer cells (human cell line) expressing green fluorescent protein (gfp)

"Natural killer cells, or NK cells, are a type of cytotoxic lymphocyte critical to the innate immune system. Typically, immune cells detect major histocompatibility complex (MHC) presented on infected cell surfaces, triggering cytokine release, causing lysis or apoptosis." [https://en.wikipedia.org/wiki/Natural_killer_cell]

GFP immunofluorescence on HEK 293 cells Icon

GFP immunofluorescence on HEK 293 cells

GFP immunofluorescence on HEK 293 cells

"Human embryonic kidney 293 cells, also often referred to as HEK 293, HEK-293, 293 cells, or less precisely as HEK cells, are a specific cell line originally derived from human embryonic kidney cellsgrown in tissue culture." [https://en.wikipedia.org/wiki/HEK_293_cells]

NK92 cells natural killer cells (human cell line) same samples microwell plate. Icon

NK92 cells natural killer cells (human cell line) same samples microwell plate.

NK92 cells natural killer cells (human cell line) same samples microwellplate

"Natural killer cells, or NK cells, are a type of cytotoxic lymphocyte critical to the innate immune system. Typically, immune cells detect major histocompatibility complex (MHC) presented on infected cell surfaces, triggering cytokine release, causing lysis or apoptosis." [https://en.wikipedia.org/wiki/Natural_killer_cell]

Mouse spleen cells mostly lymphocytes stained with FITC-conjugated anti mouse cd45 stain Icon

Mouse spleen cells mostly lymphocytes stained with FITC-conjugated anti mouse cd45 stain

Mouse spleen cells mostly lymphocytes stained with FITC-conjugated anti mouse CD45 stain

"The spleen is an organ found in virtually all vertebrates. Similar in structure to a large lymph node, it acts primarily as a blood filter. A study published in 2009 using mice found that the red pulp of the spleen forms a reservoir that contains over half of the body's monocytes." [https://en.wikipedia.org/wiki/Spleen]

Foci assay: total well view Icon

Foci assay: total well view

Foci assay: total well view

"The focus forming assay (FFA) is a variation of the plaque assay, but instead of relying on cell lysis in order to detect plaque formation, the FFA employs immunostaining techniques using fluorescently labeled antibodies specific for a viral antigen to detect infected host cells and infectious virus particles before an actual plaque is formed." [https://en.wikipedia.org/wiki/Virus_quantification]

Foci assay: Very large Foci Icon

Foci assay: Very large Foci

Foci assay: very large Foci

"The focus forming assay (FFA) is a variation of the plaque assay, but instead of relying on cell lysis in order to detect plaque formation, the FFA employs immunostaining techniques using fluorescently labeled antibodies specific for a viral antigen to detect infected host cells and infectious virus particles before an actual plaque is formed." [https://en.wikipedia.org/wiki/Virus_quantification]

Plaque assay extremely large spots Icon

Plaque assay extremely large spots

Plaque assay with extremely large spots

"The TCID50 (Median Tissue Culture Infectious Dose) is one of the methods used when verifying viral titer. TCID50 signifies the concentration at which 50% of the cells are infected when a test tube or well plate upon which cells have been cultured is inoculated with a diluted solution of viral fluid." [https://www.zeomic.co.jp/en/glossary/virus/71]

TCID50 viral assay Icon

TCID50 viral assay

CV Plaque assay.Example with large and fuzzy plaques Icon

CV Plaque assay.Example with large and fuzzy plaques

CV Plaque assay; example with large and fuzzy plaques

Cells were infected and covered with an overlay. Surviving cells stained with CV. Cells killed by the virus.

Plaque assay.Very bright plaques in 24 well plate Icon

Plaque assay.Very bright plaques in 24 well plate

Plaque assay with very bright plaques in 24 well plate

Cells were infected and covered with an overlay. Surviving cells stained with CV. Cells killed by the virus.

Neutralization assay Icon

Neutralization assay

Neutralization assay

"The plaque reduction neutralization test is used to quantify the titer of neutralizing antibody for a virus. The serum sample or solution of antibody to be tested is diluted and mixed with a viral suspension." [https://en.wikipedia.org/wiki/Plaque_reduction_neutralization_test]

Neutralization assay Accepted Plaques are surrounded Icon

Neutralization assay Accepted Plaques are surrounded

Neutralization assay

Accepted Plaques are surrounded

"The plaque reduction neutralization test is used to quantify the titer of neutralizing antibody for a virus. The serum sample or solution of antibody to be tested is diluted and mixed with a viral suspension." [https://en.wikipedia.org/wiki/Plaque_reduction_neutralization_test]

Detection of Marek's disease virus Plaques after staining with specific anti-MDV antibodies and Alexa 488 labelled secondary antibodies Icon

Detection of Marek's disease virus Plaques after staining with specific anti-MDV antibodies and Alexa 488 labelled secondary antibodies

Detection of Marek's disease virus

Plaques after staining with specific anti-MDV antibodies and Alexa 488 labelled secondary antibodies

"Marek's disease is a highly contagious viral neoplastic disease in chickens. The disease is characterized by the presence of T cell lymphoma as well as infiltration of nerves and organs by lymphocytes. Viruses related to MDV appear to be benign and can be used as vaccine strains to prevent Marek's disease." [https://en.wikipedia.org/wiki/Marek's_disease]

Detection of Marek's disease virus Plaques after staining with specific anti-MDV antibodies and Alexa 568 labelled secondary antibodies Icon

Detection of Marek's disease virus Plaques after staining with specific anti-MDV antibodies and Alexa 568 labelled secondary antibodies

Detection of Marek's disease virus but Alexa 568 labelled

"Marek's disease is a highly contagious viral neoplastic disease in chickens. The disease is characterized by the presence of T cell lymphoma as well as infiltration of nerves and organs by lymphocytes. Viruses related to MDV appear to be benign and can be used as vaccine strains to prevent Marek's disease. " [https://en.wikipedia.org/wiki/Marek's_disease]

Organoids Icon

Organoids

Organoids

"An organoid is a miniaturized and simplified version of an organ produced in vitro in three dimensions that shows realistic micro-anatomy. The technique for growing organoids has rapidly improved since the early 2010s." [https://en.wikipedia.org/wiki/Organoid]

Dual Transgene assay Icon

Dual Transgene assay

Dual Transgene assay

A segment of DNA introduced to some other organism. See [https://en.wikipedia.org/wiki/Transgene]

Various colonies in 6 well plates and Petridishes 35 mm - 100 mm Icon

Various colonies in 6 well plates and Petridishes 35 mm - 100 mm

Various colonies in 6 well plates

Various media/strain combinations on petridishes Icon

Various media/strain combinations on petridishes

Various media/strain combinations on petridishes

Antibiotic potency USP/PhEur/DIN/BP Petrishes Icon

Antibiotic potency USP/PhEur/DIN/BP Petrishes

The antibiotic potency assay measures the effectiveness of an antibiotic by the degree of growth inhibition on susceptible strains of microorganisms at different concentrations.

Adenovirus Icon

Adenovirus

Adenovirus

"Adenoviruses (members of the family Adenoviridae) are medium-sized (90–100 nm), nonenveloped (without an outer lipid bilayer) viruses with an icosahedral nucleocapsid containing a double stranded DNA genome. Their name derives from their initial isolation from human adenoids in 1953.[1] They have a broad range of vertebrate hosts; in humans, more than 50 distinct adenoviral serotypes have been found to cause a wide range of illnesses, from mild respiratory infections in young children (known as the common cold) to life-threatening multi-organ disease in people with a weakened immune system."[https://en.wikipedia.org/wiki/Adenoviridae]

SARS COV-2 Icon

SARS COV-2

In order to research and develop CoV-2 antiviral substances the ‘Inverted immunefluorescence test’ is an indispensable tool.

Example application:

Vero cells infected by SARS-CoV-2

Detected secondary Antibody: Cy3 AffiniPure  Anti -Human IgG

 

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