High Quality, Innovative Instruments for Bioscience …...Bioscience Research At Neuro Probe, Inc.,...

High Quality,

Innovative

Instruments

for Bioscience

Research

Neuro Probe, Inc.

High Quality, Innovative Instruments forBioscience Research

At Neuro Probe, Inc., we focus on helping you meet your

research goals and facilitating your important work. Our

mission is to provide high quality instruments and innovative

solutions for drug discovery and development as well as basic

research. We accomplish this through a combination of strict

quality standards, innovative products, and exemplary

customer service.

We have been designing, manufacturing, and selling

instruments for bioscience research for more than 30 years.

Since the mid-1970’s, our primary focus has been cell activity

assay instruments. We offer a unique line of products

developed specifically for the study of chemotaxis. These

products have been used as standards in the industry for

many years. We provide a wide range of high quality reusable

multi-well instruments, Boyden chambers, and the ChemoTx®

system of disposable instruments. We are currently developing

disposable instruments for high content screening (HCS) and

high throughput screening (HTS). In 2002 our product line

expanded to include a protein crystallization system.

We believe that our relationship with you is very important.

We pride ourselves on working closely with our customers

and we encourage you to contact us to discuss your projects.

Neuro Probe products are

warranted to be free from

flaws in materials and

workmanship for one year

from date of purchase unless

otherwise specified. This

warranty becomes void if

protocols for use and cleaning

of the instruments are

not followed.

Notice: Products are for

research use only. Not for use

in diagnostic procedures.

Contents

Chemotaxis Chambers and Accessories

ChemoTx® Disposable System........................................................4

Reusable 96-Well Microplate Chambers ......................................6

Reusable 96-Well Chambers..............................................................8

10, 12, and 48-Well Chambers .........................................................9

Specialty Chambers .............................................................................11

48-Well Chamber with Filter Retaining Hardware...........11

Three-Tiered Chamber.................................................................12

Cell Orientation Chamber (Zigmond) ..................................12

Single-Well Chambers .........................................................................13

Filter Information..................................................................................14

Protein Crystallography

Cell Settling & Differential Counting

Price List (back pocket)

3

4

15

16


Neuro Probe ChemoTx® DisposableChemotaxis System101, 106, 111, and 116 Series

The Leading Edge in Speed and Convenience

The ChemoTx® System is a disposable cell migration system. Basedon the standard 96-well microplate format, it is compatible with most96-well fluorescent and densitometric plate readers, as well as mostliquid-dispensing robots.

The system consists of all the chamber components necessary toperform a basic chemotaxis assay: a microplate, filter, and lid. Platesare molded from tissue culture grade transparent polystyrene, with30µL or 300µL well capacity.

The filters for this system are bonded to frames for ease of handlingand selectively coated with a hydrophobic mask around each of thetest sites. The hydrophobic mask restricts the cell suspension to thesesites, eliminating the need for upper wells. Test sites are available intwo exposed filter areas, 8mm2 or 25mm2. Additional hydrophobiccoatings, combined with the proprietary design of the plates, providethe seal between the bottom of the filter and the rims of the plate,so there is no need for gaskets and clamping hardware. Bubble

entrapment is eliminated in both the top and bottom fluidsby this design. The lid aids in reducing evaporation

and facilitates stacking.

There are four possible combinations ofwell volume and exposed filter area.

For most fluorescence assays the101 series, with 30µL well

volume and 8mm2 filter

4

area, works well. If the number of migrated cells does not createsufficient signal strength relative to the background, the 106 series,with 25mm2 area, will yield increased signal strength. For incubationtimes longer than 3–4 hours, the larger, 300µL plate volume willallow for more media to sustain the cells. When quantifying with adensitometric reader, the 116 series will provide a larger exposedfilter area for increased signal strength and deeper wells to simplifythe staining process and enhance reading. If using the 300µL wellvolume is not desirable, e.g., when using rare or expensive reagents,an alternative is to use the 106 series with 30µL well volume for themigration and then use the 96-well funnel plate to transfer themigrated cells into a deeper plate for staining and counting.

Polycarbonate membranes are available in a wide range of poresizes. Please see the price list or visit our website for a full listingof standard configurations and pore sizes available. Informationregarding pore density as it relates to pore size may be found onthe “Filters/Membranes” page.

5

Accessory

96-Well Funnel Plate(Stock# FP1). Allows rapidtransfer of cells and mediafrom the 30µL plate intoother 96-well plates.

101 Series

106 Series

116 Series

866.417.0014 Tel 301.977.5711 Fax NeuroProbe.com


Neuro Probe MB Series Microplate Chambers MBA96, MBB96, & MBC96

Speed And Objectivity Combined with Versatility

The reusable MB series chemotaxis chambers are designed forchemotaxis assays, tumor cell invasion assays, and cell-cell interactionstudies. They use Neuro Probe framed filters and select 96-wellmicroplates. The plates are compatible with standard automatedreaders. Use of automated readers has two significant advantagesover visually counting cells. The first is speed and the second isobjectivity; viewer fatigue is eliminated. These advantages apply toall Neuro Probe 96-well chambers, however this design utilizes aremovable plate as the lower wells, making it convenient for assaysinvolving non-adherent cells. The MB series chambers maintain allthe capabilities of the rest of our reusable chambers, including workwith adherent cells.

Each chamber consists of precision-machined top and bottomplates, hinged together, enabling rapid and accurate assembly of thecomponents. The top plate has 96 through holes; the bottom platehas a single large recess sized to hold a microplate. This microplateforms the lower wells. A combination of a spacer and a siliconegasket ensures a seal around each well.

A framed filter is placed over the microplate.Filters with adhesive are available to bondto microplates, forming units that can becentrifuged. Standard format fluores-cence, densitometric, scintillation, andluminescence readers can be used tocount migrated cells in the microplate.Many fluorescence readers can also beused to count cells on the filter. Please seethe “Filters/Membranes” page for filter specifi-cations, and refer to the “Framed Filters” section of the price listor visit our website for available pore sizes and types.

The standard plates (stock# MPC) have a well capacity of 400µL,and may be used in any of the MB series chambers. Special lowvolume plates (stock# MP30) may also be used in the MBB96 orMBC96 chambers, with the purchase of an appropriate spacer(stock# SP30). An additional advantage to using these low volumeplates is that filters with adhesive are not needed, as the filter andplate press together. Reusable 96-well inserts may be used in place ofdisposable microplates. These inserts can also be used in automatedreaders. The optional inserts are listed in the table on the next page.

6

MBA96

MBB96

MBC96

866.417.0014 Tel 301.977.5711 Fax NeuroProbe.com 7

MP30 plate with SP30 spacer, MPC plate with SP3 spacer

Low-volume MB inserts with spacers

The three MB series chambers differ in upper well volume andexposed filter area. Lower well volume is determined by themicroplate selected.

Stock# For Use With Well area Volume

IA-100 MBA96 32mm2 100µL

IB-50 MBB96 18mm2 50µL

IC-25 MBC96 8mm2 25µL

Each reusable insert includes the required spacer.

Stock# Upper Well Exposed Filter Well DiameterCapacity (µL) Area (mm2) (mm)

MBA96 395 32 6.4

MBB96 225 18 4.8

MBC96 50/225* 8 3.2

*The MBC96 has a step in the upper well: capacity is 50µL when filled tothe step, or 225µL if filled to the top.

Replacement gaskets and spacers are available. Please refer to the pricelist or visit our website for more information. 96-well funnel plates(Stock# FP1) may be used to transfer fluids from the MP30 plates, orthe IB-50 or IC-25 reusable inserts, into other 96-well plates.


Neuro Probe A Series 96-Well ChambersAA96, AB96, & AC96

Speed and Objectivity in a Value Series

The A series 96-well chemotaxis chambers are reusable instrumentsconsisting of precision-machined top and bottom acrylic plates, asilicone gasket, and assembly hardware. The bottom plate has 96wells; corresponding holes in the top plate form the upper wells.Registration pins ensure easy and accurate assembly. The gasket anda framed filter can be installed on the top plate prior to chamberassembly. High quality finishes on the interior surfaces facilitatecleaning and reduce the possibility of contamination.

After incubation, adherent cells tagged with a fluorescent marker maybe quantified by fluorescence-type readers. Alternatively, the cellsmay be stained and counted with a microscope.

The three A series chambers differ in well volume and exposed filterarea as follows:

8

AA96

AB96

AC96

Stock# Exposed Lower Well Upper Well Filter Area Capacity Capacity

AA96 32 mm2 80µL 390µL

AB96 18 mm2 30µL 225µL

AC96 8 mm2 25µL 50/225µL *

*The AC96 has a step in the upper well: capacity is 50µL when filled tothe step, or 225µL if filled to the top.

For additional information regarding framed filters, please see the“Filters/Membranes” page, and refer to the “Framed Filters” section ofthe price list or visit our website for available pore sizes. Replacementgaskets (A96G, B96G or C96G) are available, as are thumb nuts(N8L). No accessories are required, although the filter wiper (P48W)may be used to remove non-migrated cells from the top of the filterbefore counting.


The Neuro Probe Standard Chemotaxis SeriesAP48, AA12, & AA10

Proven Dependability for Cell Migration

The AP48 “microchemotaxis chamber” has long been the standard inthe field of cell migration. After migrating, cells on the membrane arecounted visually with a microscope. The AP48, AA12, and AA10chambers consist of two precision-machined acrylic plates, a siliconegasket, and hardware for assembly. High quality finishes on the interiorsurfaces facilitate cleaning and reduce the possibility of contamination.

9

AP48

AA10

AA12

These chambers may be used with polycarbonate or cellulose nitratefilters. They use one piece of membrane, 25x80mm, to cover all thewells. Please see the “Filters/Membranes” page for more informationand refer to the price list or visit our website for availability of poresizes.

The 48-well chemotaxis chamber (stock# AP48) has a lower wellvolume of 25µL. Corresponding holes in the top plate form theupper wells, with volumes of 50µL. The membrane is placed overthe filled lower wells, then the gasket and top plate are added andfastened down to create a seal between the two plates. Each well hasan exposed filter area of 8mm2. The wells are arranged in groups ofsix to facilitate visual orientation and pattern recognition. This well-proven design is still an excellent choice for many experiments.

Neuro Probe 12-well or 10-well chambers are designed for studyinga small number of samples, or when working with larger volumesthan the AP48 allows. The 12-well (stock# AA12) and 10-well(stock# AA10) chemotaxis chambers have bottom plates with tworows of six or five wells, respectively. Volumes are 150µL for theAA12 and 400µL for the AA10. Corresponding holes in the top


10

P48AP

plate form the upper wells, with volumes of 100µL for the 12-welland 285µL for the 10-well. As with the AP48, the membrane isplaced over the filled lower wells, then the gasket and top plate areadded and fastened down before adding cells to the upper wells.Each well has an exposed filter area of 18mm2 (AA12) or 50mm2

(AA10).

Replacement gaskets and hardware are available for all reusablechambers (please see the price list or visit our website for stocknumbers). We also offer a range of accessories: a filter wiper (P48W)suitable for mounting on a 1/2" or 12mm ring stand, large and smallclamps (P48CL and P48CS, respectively), and curved stainless steelforceps (P48F). These may be purchased individually, or in a kit

(P48AP) which includes one each of thewiper and large and small clamps and twopairs of forceps. A template (P48TM) isavailable for the AP48, which can assist inlocating migrated cells on the polycarbon-ate filters if needed.

P48TM


Specialty ChambersNeuro Probe Specialty Chambers cover a variety of applications.If you do not see what you need here, please call or e-mail our CustomerService Department to discuss your requirements. We can often modifyone of our standard products to suit other applications.

Special Neuro Probe 48-WellMicrochemotaxis Chamber AC48

For Use with Cellulose Nitrate Membranes

The 48-well chemotaxis chamber with filter retaining hardware(stock# AC48) is similar to the standard chamber (AP48), but hasbeen modified to facilitate its use with cellulose nitrate membranes.It has eight screws mounted in the bottom surface of the top plate.Holes in the gasket snap over these screw heads, allowing the filtersto be inserted between the gasket and screw heads. These items maythen be lowered as a unit onto the bottom plate.

The bottom plate has 48 wells, each with 25µL volume. Matchingholes in the top plate form the upper wells, with volumes of 50µL.Two pieces of membrane are used, each 35×41mm. Each well hasan exposed filter area of 8mm2.

The wells are arranged in groups of six, similar to the standard48-well chamber. This facilitates visual orientation and pattern recog-nition, but the spacing is farther apart than on the AP48 to reducethe possibility of crosstalk during extended incubations with cellulosenitrate filters. This chamber may also be used with polycarbonatefilters, but if that is your primary interest, you may also want toconsider the more commonly used AP48 Chamber. Please see the“Filters/Membranes” page for more information on filter types andrefer to the price list or visit our website for availability of pore sizes.

Replacement gaskets (C48G) and hardware are available for thischamber. A set of four stainless steel clips (Stock# SSC4) is availablefor mounting cellulose nitrate filters on microscope slides. We alsooffer a range of accessories for working with polycarbonate mem-branes: a filter wiper (P48W) suitable for mounting on a 1/2" or12mm ring stand, large and small clamps (P48CL and P48CS,respectively), and curved stainless steel forceps (P48F). These may bepurchased individually, or in a kit (P48AP)that includes one wiper, one large clamp,one small clamp, and two forceps. A tem-plate (C48TM) is available for the AC48chamber, which can assist in locatingmigrated cells on polycarbonate filtersif needed.

11

AC48

AC48 disassembled

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

C48TM and SSC4


Neuro Probe Three-Tiered ChamberA3BP48

Modification of AP48 for Use withInverted Microscope

The Neuro Probe Three-Tiered Chamber (Stock# A3BP48) is aspecial modification of the standard AP48 chamber. This chamberfeatures flat bottom wells. This configuration facilitates the observa-tion of cells on the bottom of the filter without disassembling thechamber. The chamber consists of three plates. The bottom plateis clear with no wells, and the top and middle plates both have48 through holes with volumes of 50µL. Special hardware that holdsall three pieces together is permanently mounted in the middle plate.The chamber is sealed using two gaskets and two sets of thumbnuts.

A second filter may be placed between the middle and bottomplates, allowing cells that do not adhere to the underside of theprimary filter to settle out onto this secondary filter for counting.Prior to the development of the 96-well microplate chambers, thiswas the simplest means for working with non-adherent cells. Thischamber is compatible with the same accessories as the AP48.Replacement gaskets and hardware are also available.

The Neuro Probe Zigmond ChamberZ02

For High-Resolution Morphologic Studies

The Zigmond chamber (Stock# Z02) is designed for morphologicstudies of cell migration. High-resolution light microscopes may beused to view cells as they orient and migrate. The chamber is madefrom a 2mm thick, 25×75mm glass microscope slide. It has twogrooves 1mm deep separated by a 1mm wide section referred to asthe “bridge.” There is a hole at each end of the slide to accept a pinclamp. These clamps are used to hold a 22×40mm cover glass inplace during incubation and viewing of the cells. No other accessoriesare needed.

12

A3BP48

A3BP48 disassembled

Z02

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .


Neuro Probe Single-Well ChemotaxisChambersBW25, BW100, BW200S, BW200L, & BY312

Blind Well and Boyden Chambers

The term “Boyden Chamber” has become a generic label for anyinstrument used to observe cell migration. Neuro Probe’s BoydenChamber (Stock# BY312) has a U-shaped lower well open to theatmosphere, thus equalizing pressure across the membrane. The blindwell chambers have a lower well with a hemispherical bottom thatis completely enclosed, or “blind,” except where the membraneseparates it from the upper well.

Blind well and Boyden chambers are precision-machined from clearacrylic. The high quality finishes on both interior and exterior surfacesfacilitate cleaning, reduce the chances of contamination, and permitdetection of bubbles trapped below the filter. Filter retainers madeof white acetal are included with the chambers. The retainer formsa seal without requiring a gasket, reducing setup and cleaning time.These chambers all use 13mm diameter filters, sold separately. Pleaserefer to the “Filters/Membranes” page for more information and referto the price list or visit our website for availability of pore sizes.

The following table shows the various configurations available, withvolumes and exposed filter areas.

Use of the blind well chambers can be advantageous in small samplenumber studies. The need to perform parallel assays at least in dupli-cate, with corresponding positive and negative controls, requires theuse of, at minimum, four chambers, and preferably six or more. Weoffer replacement filter retainers for all of our single well chambers.Please see the price list or visit our website for specific stock numbers.We also offer a blindwell rack (Stock# BWR24) that will hold up totwenty-four blind well chambers, and facilitates moving multiplechambers into and out of the incubator. However, if you areperforming larger scale studies, you may want to consider one of ourmulti-well chambers. Please contact Customer Service for assistance.

13

BW25, BW100, BW200S, BW200L and BY312

BWR24Stock # Exposed Filter Lower Well Upper Well

Area (mm2) Volume (µL) Volume (µL)

BW25 18 25 200

BW100 18 100 200

BW200S 18 200 200

BW200L 50 200 800

BY312 50 1500 560


Filters/Membranes for Chemotaxis

A Large Selection to Suit Your Applications

Polycarbonate track-etch (PCTE) membranes are the primary type offilters used in our chambers and instrument systems. This is a verythin membrane ranging between 6–10µm in nominal thickness. Thismembrane type offers excellent uniformity in pore size and density,as well as other benefits such as low extractables, low protein binding,and negligible absorption. These characteristics make it ideally suitedfor use in chemotaxis assays as well as other high purity screening assays.

PCTE membranes are available pre-treated with the wetting agentpolyvinylpyrrolidone (PVP) which makes the surface hydrophilic.PVP-treated membranes are used when the media solutions cannotwet out the membrane by themselves. This occurs primarily whenworking with very small pore sizes or high viscosity media solutions.Certain cell types, neutrophils for example, and extra-cellular matrixproteins do not adhere well to PVP-treated membranes. A carefulassessment should be done before selecting this membrane treatment.

Many factors other than cell type enter into the pore size selectionprocess. Please visit our website (www.neuroprobe.com), and use thesite’s search function to find more information on Experiment Design.Also see the web site or refer to our price list for pore size availabilityfor specific products.

The ChemoTx® disposable system and the reusable 96-well cham-bers use relatively large pieces of PCTE membrane that are difficultto handle by themselves. For convenient handling, these instrumentsuse filters bonded to anodized aluminum frames, referred to through-out the catalog as “framed filters.” Framed filters with adhesive(Stock# ending in “/A”) are available for use in the MB series cham-bers only. The adhesive makes it possible to remove the microplatefrom the chamber with the filter still attached for spinning in a centrifuge.

The Standard and Specialty series of multiwell chambers and thesingle well chambers use punched filters in a variety of sizes. Single-well chambers, both blind well and Boyden, use 13mm diameterfilters. Most of the multiwell chambers use 25×80mm filters, theexception being the AC48 that uses 35×41mm filters.

Filters for the Standard and Specialty series are also available incellulose nitrate. These are much thicker (~150mm) fibrous filters,sometimes referred to as “torturous path” filters. Rather than countingcells that have migrated through the membrane, as is commonly donewith PCTE membrane, these filters are clarified with solvents afterincubation, and a high-resolution light microscope is used to observehow far into the filter the cells have migrated. This is also known as a“leading edge” study.

14

Pore size (+0/–20%) 2µm 3µm 5µm 8µm 10µm 12µm 14µm

Density (pores/mm2) 2 × 104 2 × 104 4 × 103 1 × 103 1 × 103 1 × 103 5 × 102

(tolerance +/– 15%)

Polycarbonate membranes are available inthe following pore sizes and correspondingpore densities:


Neuro Probe Protein CrystallographyCT300L2/5

A 96-Well System for the Hanging Drop Method

The Neuro Probe crystal plate system (Stock# CT300L2/5) is adisposable instrument in the standard 96-well format that uses theproven hanging drop method. It consists of three components: a clearpolystyrene microplate that is pre-greased for easy sealing, a singlecover film with printed well positions for positive location, and a lidto protect the cover film and facilitate stacking. No other componentsare required. Other features include:

� Small drop volume� Easy crystal retrieval� Excellent imaging qualities� Fast setup� Saves space in the cold room (compared with 24-well plates)� May be used manually or automated with select robots, e.g., the

Gilson 925 PC Workstation

15

Protein Crystallography

CT300L2/5

ungreasedmicroplate

CT300L2/5pre-greased microplate

Photo courtesy of GlaxoSmithKline

Cell Settling & Differential Counting

Neuro Probe Cell Settling Chamber andAccessoriesDC8, DC8WF1, DC8SC, DC8AP

For Differential Counting of Cells

The cell settling chamber (Stock# DC8) is designed to deposit cellsin a monolayer on a glass microscope slide. After the wicking andsettling process, cells adhere to the slide in a random distributionsuitable for differential counting, with excellent cellular morphology.

The chamber consists of two precision-machined acrylic plates, asilicone gasket, and assembly hardware. The lower plate has a recessto fit a 25×75mm glass microscope slide. The upper plate has eightthrough holes, which form 280µL wells when the upper plate isfastened onto the lower plate with a microscope slide in place. Thegasket and a sheet of prepunched filter paper (Stock# DC8WF1)pinched between the plates allow liquid in the cell suspension to wickaway as the cells settle onto the slide, leaving the cell sites dry.

For best results, microscope slides with enhanced binding propertiesand a hydrophobic mask around the eight settling sites (Stock#DC8SC) are available. These slides improve cell adherence and alsorestrict pipetted solutions to individual sites. Different solutions canthen be applied to settled cells without crosstalk.

When working with cells of relatively low density, such as lympho-cytes, the chamber should be spun in a centrifuge equipped witha microplate carrier. A centrifuge accessory pack (Stock# DC8AP)provides adapters and a counterweight for this purpose

16

Accessories for the DC8 Chamber

DC8

Neuro Probe, Inc.

16008 Industrial Drive

Gaithersburg, MD 20877

USA

866.417.0014 Toll Free

301.417.0014 Tel

301.977.5711 Fax

www.neuroprobe.com

Distributors:

Ieda Trading Corp.

3-14-16 Hongou

Bunkyo-ku Tokyo 113

Japan

3-5804-4733 Tel

3-3816-2917 Fax

E-mail: [email protected]

Receptor Technologies, Ltd.

The Barn, Croft Lane

Adderbury, Oxon OX17 3NB

United Kingdom

(0) 1295 812600 Tel

(0) 1295 812700 Fax

E-mail: [email protected]

To receive notification of new product releases,

please send an email to [email protected]

or register on our website.

All specifications and product information is

subject to change without notice.

© 2006 Neuro Probe, Inc.

High Quality, Innovative Instruments for Bioscience …...Bioscience Research At Neuro Probe, Inc.,...

Documents

Transcript of High Quality, Innovative Instruments for Bioscience …...Bioscience Research At Neuro Probe, Inc.,...