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Cellular imaging made easy

A streamlined workflow for cell counting and phenotypic characterization is critical to many experiments. Our
SpectraMax i3x microplate reader with MiniMax cytometer provides you with cellular analysis capability in a small
footprint without the need for a complex imaging system. It eliminates the need for cell staining with StainFree Cell
Detection Technology. Combined with the analysis capabilities of SoftMax Pro Software, we provide quantitation in
an easy-to-read, user-customizable report format.

eBook contents
Image and analyze slide-based samples............................ 2 Detect cell apoptosis................................................................. 6
Measure neurite outgrowth..................................................... 3 Optimize GFP transfection....................................................... 7
Analyze cell confluence............................................................ 4 Image whole organisms and tissues.................................... 8
Alternatives to DAPI staining................................................... 5 System configurations............................................................... 9

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Image and analyze slide-based samples
Biological samples ranging from bacterial smears to tissue cross sections are
commonly studied on microscope slides. Using bright-field and fluorescence
imaging, researchers can study cytological or morphological changes
in their slide-based samples. Slide microscopy has traditionally required Identify and analyze a variety of
microscope systems, but now microplate reader systems can perform slide- different tissue types
based imaging. Here we demonstrate how the SpectraMax i3x Multi-Mode Create custom plate dimensions for
Microplate Reader with SpectraMax MiniMax 300 Imaging Cytometer can microscope slides, petri dishes and
acquire high-quality images and data from slide-based samples. other non-microplate formats
Accessible to non-imaging experts
Download Application Note with intuitive SoftMax Pro Software

A B

C D

Custom plate dimensions for the slide adapter. A custom plate setting Image analysis of FluoCells prepared slide. A pre-made slide containing
was created in SoftMax Pro Software to accommodate the slide adapter BPAE1 cells stained with Alexa Fluor 488 Phalloidin and MitoTracker Red
using the dimensions shown above. CMXRos was imaged using the MiniMax cytometer. Green fluorescence
channel images (A) and red fluorescence channel images (B) were overlaid
(C), and cell confluency was determined by Field Analysis of the green
fluorescence channel (D).

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Measure neurite outgrowth
Neurons create connections via extensions of their cellular body, which is
referred to as neurite outgrowth and is regulated by complex intracellular
signaling events. Understanding the signaling mechanisms driving neurite Acquire high-quality neuronal images
outgrowth provides valuable insight for interpreting neurotoxic compound Accelerate time to results using
screening data and for elucidating factors influencing neural regeneration. MetaMorph softwares automated
The SpectraMax MiniMax cytometer and MetaMorph software provide a image processing and analysis tools
total solution to elucidate the mechanisms behind neuron differentiation and Automatically generate detailed
regeneration and interpret neurotoxic screening results. neurite outgrowth data such as
number of processes, process
Download Application Note length, and total measured
outgrowth

Neuronal cell image. Neurons were imaged using the MiniMax cytometers
green fluorescence channel. This image represents one of twelve sites
imaged in the well.

Neuron image montage. Raw images were acquired using the MiniMax MetaMorph software object overlay. The MetaMorph softwares neurite
cytometer and stitched together using MetaMorph software. This image outgrowth module used the grayscaled neuronal image montage (top) to
montage was then used for neurite outgrowth analysis. identify neuronal cell bodies and their processes, which are indicated by a
red image overlay (bottom).

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Analyze cell confluence
Imaging cell-based assays typically requires the use of fluorescent probes
that can be toxic to living cells or may only function in fixed cells. A label-free
method for analyzing cell counts and cell confluence enables researchers
to quantitatively monitor cell proliferation and health without time-consuming Count cells and estimate confluence
workflows that may disrupt cell viability. The SpectraMax i3 reader with without fluorescent dyes
MiniMax cytometer uses StainFree Cell Detection Technology allowing you
Monitor cell growth without
to perform cell proliferation, cytotoxicity, and other assays without nuclear disrupting cells
stains like DAPI, which intercalates with DNA, or live cell dyes that are toxic
Set up imaging assays quickly and
to cells in the long term.
easily with SoftMax Pro Software
Download Application Note

StainFree analysis in SoftMax Pro Software. Left: To create a new StainFree analysis setting, the
mouse is used to draw on the image, indicating individual cells (yellow) or non-cellular areas (blue).
Right: A purple mask shows the objects identified in the image.
Cell count

Cells seeded per well

StainFree technology compared to fluorescence cell counting. Cells counted using StainFree
technology (blue), red nuclear stain (red), and green whole-cell stain (green). Counts obtained from all
three methods agree closely, demonstrating that StainFree technology gives accurate cell counts while
eliminating the need for fluorescent dyes (R2 > 0.99 for each plot).

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Alternatives to DAPI staining
DAPI (4, 6-diamidino-2-phenylindole) is a fluorescent dye often used to stain
nuclear DNA. It is employed in imaging experiments such as fluorescent
microscopy, chromosome spreads, FACS, and cell-based assays. However,
excitation with UV results in a photoconversion of DAPI that leads to Enable live cell counting while
detection of DAPI fluorescence in the FITC/GFP channel of an imaging maximizing cell viability with
StainFree technology
system, causing errors in interpretation of results. DAPI also requires cells
to be fixed for maximal staining. Here we show some alternatives to DAPI Eliminate fixation steps required prior
staining, including StainFree technology on the SpectraMax i3 reader with to staining
MiniMax cytometer, which requires no staining at all and can be used with Save time and money by avoiding
live cells. costly staining procedures

Download Application Note

StainFree Live Red Dye DAPI

Image Stain (30 min) Fix (30 min)

Image Wash (5 min)

Stain (30 min)

Wash (5 min)
Cell counts

Image

Workflow for cell analysis with StainFree Technology vs. Live Red Dye
vs. DAPI. The StainFree workflow saves about 70 minutes compared to
fixation and staining with DAPI. Moreover, cells analyzed using StainFree
Initial seeding density (cells/well)
technology remain fully viable and can be used in additional assays.

Cells counted with the MiniMax cytometer. Cells stained with Live Red
Dye were imaged in the red fluorescent (top left) or transmitted light (top
middle) channel. StainFree cell counts (top right, purple masks) correlate
very closely to cell counts based on red nuclear staining, as shown in the
graph (green circles, StainFree counts; red circles, red nuclear counts). The
r2 values for both curves were 0.99.

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Detect cell apoptosis
Tumor necrosis factor (TNF) is one of the key inflammatory cytokines,
which modulate various events in several cellular pathways. It is known
to induce apoptosis in hematopoietic cells, such as U937, via activation of Homogeneous assay with easy,
intracellular caspase cascades. Granulocyte-macrophage colony-stimulating mix-and-read workflow
factor (GM-CSF), on the other hand, is a member of hematopoietic growth
Optimized for fluorescence
factors that promotes proliferation and/or differentiation. However, it has microplate readers
been reported that U937 cells show growth inhibition and apoptosis in
Increased throughput with
response to GM-CSF. While both TNF and GM-CSF induce apoptosis in
microplate reader format
U937 cells, the time course appeared to be distinct between these two
cytokines. When combined, a highly synergistic effect of the two cytokines Preconfigured protocol in SoftMax
was observed. In this cell-based study, we used the EarlyTox Cell Viability Pro Software
Assay Kits, a family of reagents for assessing cell viability and various
apoptotic events, to measure several cellular readouts on the SpectraMax
i3x reader with MiniMax cytometer.

Download Application Note

EarlyTox Cell Viability Assay Kits Webpage

Healthy cells
(can be detected
with Live Cell Kit)

Cytokine treatment for 24 or 48 hours

Add Add Add


Live/Dead reagent Caspase-3/7 R110 reagent NucView 488 reagent

Detect cell viability Detect cell apoptosis in Detect cell apoptosis in


E ndpoint readout on cell lysate live cells
microplate readers Endpoint readout on Long time course
microplate readers readout on microplate
readers or cytometer
Evaluation of GM-CSF and TNF-induced apoptosis with EarlyTox Cell Viability Assay Kits.

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Optimize GFP transfection
Expression of fluorescent proteins, for example, green fluorescent protein
(GFP), is often used as a readout for reporter gene assays. In a typical
reporter assay, the regulatory sequence of a gene of interest is attached Identify transfected vs.
to a reporter gene that can be easily detected and measured. With a non-transfected cells easily
GFP reporter, increased activity of the gene of interest leads to increased
Obtain quantitative data on relative
expression of fluorescent protein in the cell, which can be measured on a transfection efficiencies accurately
fluorescence microplate reader or imaging cytometer. Here we demonstrate
Measure cell confluence prior to
how the SpectraMax i3 reader and MiniMax cytometer are used to monitor
transfection
transfection efficiency and speed the time to results for assays where
transfection is key to success. Faster time to results for reporter
assays with SoftMax Pro Software
Download Application Note

Transmitted light GFP fluorescence

A B
% confluent at transfection

0.25 L reagent
0.1 g DNA +

C D
Cells seeded per well
0.75 L reagent
0.2 g DNA +

Cell confluence. Percent confluence of cells just prior to transfection was


determined using the imaging cytometer. For initial seeding densities of
15000, 10000, and 5000 cells per well, the measured percent confluence
was 44, 30, and 15, respectively.

Imaging of transfected cells. Transfected cells were imaged using


the TL (A, C) and green fluorescent (B, D) channels of the SpectraMax
MiniMax cytometer. Cells shown were seeded at 15000 cells per well.
Two transfection conditions are shown, representing low and high
transfection efficiencies.

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Image whole organisms and tissues
Across labscapes, creatures of all sorts come into focus. Enjoy this gallery of the beautiful creatures (and parts thereof) weve
imaged with the SpectraMax MiniMax cytometer.

View Webpage

The nematode C. elegans has long served as Heart muscle with H&E staining, imaged in the
a model system for developmental biology TL channel of the MiniMax cytometer.
due to its simplicity and fully characterized
pattern of cell lineages. Here, C. elegans Representative section of rat brain from a
expressing GFP were imaged using the slide imaged with the 541 nm fluorescence
transmitted light and green fluorescent channels channel of the MiniMax cytometer.
of the MiniMax cytometer.

Located in the mammalian cochlea, the


organ of Corti transduces sound vibration
into nerve signals. Here, organs of Corti with
GFP-expressing cells were isolated from mouse
and imaged in the transmitted light and green
fluorescent channels of the MiniMax cytometer.
Inset, enlarged detail of cellular layers.
Zebrafish are heroes of the vertebrate Auto-fluorescence of Arabidopsis thaliana
genetics world. They are amenable to mutant seeds imaged in the 541 channel of the
screens and small enough to fit into the wells MiniMax cytometer.
of a microplate. If you can get them to slow
down, you can get beautiful images like these.
Here, a GFP-expressing zebrafish was stained
with AlexaFluor 647-labeled phalloidin, which
highlights its tail muscles.

Jumping and running are brought to you


by the gastrocnemius muscle. Here, mouse
gastrocnemius muscle was stained with a
muscle-specific antibody and imaged on the
transmitted light (left) and 541 nm fluorescence
(right) channels of the MiniMax cytometer.

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System configurations

Technical specifications for MiniMax cytometer


Proprietary solid state illumination,
Light source
White, 460/20 nm and 625/20 nm excitation
Detector 1.25 megapixel, 12-bit high sensitivity CCD camera
Emission Brightfield; Green 541/108 nm; Red 713/123 nm
Objective Single 4X objective
Autofocus Proprietary laser scanning autofocus
Resolution 1.9 m x 1.9 m pixel size
Data acquisition and
SpectraMax i3x microplate analysis software
SoftMax Pro Software MiniMax Imaging Edition

reader with MiniMax Imaging Speed* Acquisition Acquisition + Analysis


cytometer 1 color 96 wells 3:40 6:30
2 color 96 wells 3:40 6:30
View Video Specimen carriers ANSI/SBS-conformant microplates, 96- and 384-wells
39.2 cm W x 19.5 cm H x 60.6 cm D (SpectraMax MiniMax 300 Imaging
Download Data Sheet Dimensions (cm)
Cytometer), 39.2 W x 44.0 H x 60.6 L (with base system)

Request Quote * Using single site acquisition with 10 ms exposure time.

Products Part number


SpectraMax MiniMax 300 Imaging Cytometer 5024062
SpectraMax MiniMax 300 Desktop Computer 5029422
SpectraMax MiniMax 300 22 Monitor 5024296
SpectraMax i3x Multi-Mode Microplate Reader i3x

Designed specifically for our instruments, our assay kits are optimized for
maximum performance and are supported with software protocols and analysis
that enable you to go from samples to answers quickly.

View Assay Kits

Contact Us
Phone: +1-800-635-5577
Web: www.moleculardevices.com
Email: info@moldev.com
Check our website for a current listing
of worldwide distributors. 9

The trademarks used herein are the property of Molecular Devices, LLC or their respective owners. 2017 Molecular Devices, LLC
Specifications subject to change without notice. Patents: www.moleculardevices.com/productpatents 5/17 2107A
FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES. Printed in USA

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