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3556 HEMACYTOMETER / 3557 NEUBAUER COUNTING CHAMBER: OPERATION INSTRUCTIONS

SAVE THESE INSTRUCTIONS FOR FUTURE USE

Info: The most current version of these instructions can be found online at www.grahamfield.com.

WARNING: Important! Read and understand these instructions before operating the Hemacytometer. If the Hemacytometer is not properly assembled, personal injury and damage to the Hemacytometer could result.

WARNING: If components are damaged or missing, contact your GF authorized distributor immediately. DO NOT use substitute parts. Use only Grafco replacement parts. Non-Grafco replacement parts could cause personal injury and damage to the Hemacytometer.

WARNING: Notice for California Customers- California Proposition 65 WARNING: This product contains a chemical known to the State of California to cause cancer and reproductive or developmental harm.

WARNING: For use with rubber suction bulbs or pipette pumps. DO NOT pipette by mouth.

WARNING: GF Health Products, Inc. assumes no responsibility for any damage or injury caused by improper assembly, installation or use of this product.

PRODUCT DESCRIPTION

Counting Chamber (Hemacytometer) Intended Use

A counting chamber is a precision measuring instrument made of special optical glass. It is used to count cells or other particles in suspension under a microscope.

Counting chambers are mainly used for blood analysis (counting leucocytes, erythrocytes and thrombocytes) and to count cells of liquor. Counting chambers are also used to count bacteria and fungus spores.

COMPONENTS INCLUDED

3556: Complete Hemacytometer Set includes

1 ea Double Improved Neubauer Counting Chamber1 ea Leucocyte Pipette (white bulb)1 ea Erythrocyte Pipette (red bulb)2 ea 16 cm transparent silicone tubing (color coded for each pipette)2 ea 0.4 mm glass coverslip in a clear plastic protective case

3557: Double Improved Neubauer Counting Chamber includes

1 ea Double improved Neubauer counting chamber2 ea 0.4 mm thick, 20 mm W x 26 mm H glass coverslips individually

packed in a clear plastic protective case

3556 Hemacytometer

tubing5.5 mm OD x 3.5 mm IDx 16 cm long

tubing red end(for use withErythrocyte pipette)

tubing white end(for use with

Leucocyte pipette)

Erythrocyte pipette(red bulb)

countingchamber

Leucocyte pipette(white bulb)

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COUNTING CHAMBER DESCRIPTION

There are four widthwise grooves in the central third of a rectangular, thick base plate made of special optical glass, shown above. The grooves are parallel to the short sides of the base plate and the central third has the same size as the coverglass used with the counting chamber. The two larger external surfaces are unfinished and are used for marking purposes.

The central support and the two external supports are ground smooth and polished. The surface of the central support is deeper than that of the two external supports. The counting nets are engraved in the central support (chamber base).

When a cover glass is placed on the external supports, a capillary gap is produced between the underside of this cover glass and the central support of the counting chamber, as shown above.

This counting chamber has double net ruling, shown above: middle support with one division (two counting nets).

Each counting net is made up of a square net division which is not visible until it is placed under a microscope (approximately 100 times magnification).

Counting Net Dimensions

Largest square size : 1 mm²

Group square : 0,04 mm²

Smallest square size : 0,025 mm²

Chamber Depth is 0.100 mm

The net division of these chambers has 3 times 3 large squares, each with an area of 1 mm².

The Four Corner Squares are Used for Leucocyte Counts

The large square in the middle is also divided into five times five group squares with an edge length of 0.2 mm each and an area of 0.04 mm2 each. The group squares in turn are divided into sixteen very small squares with an area of 0.0025 mm2 each.

Five of These Group Squares are Used for Erythrocyte Counts

Special attention should be given to the fact that the chamber has triple lines on all sides, of which the central line is to be regarded as the actual dimension line. This is important for deciding whether cells in the border area are to be counted or not.

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HOW TO FILL THE COUNTING CHAMBER

Sliding on the Cover Glass

Moisten the external supports with distilled water, then push the cover glass gently onto the counting chamber from the front, as shown above.

s NOTICE: The cover glass is fragile! Take care when handling.

The formation of interference lines (Newton rings) between the external support and the cover glass shows that the cover glass is correctly positioned.

Feeding

Take a well mixed pipette from the shaker and dispose of the first few drops.

Wipe the pipette dry on the outside and then hold it at an angle until a small drop has formed at the tip of the pipette, as shown above. This drop is then to be placed between the cover glass and the counting chamber.

As a result of the capillary effect the gap between the cover glass and the chamber base fills up.

Before the thinned blood solution can overflow at the edges of the chamber section, the tip of the pipette must be removed. If any air bubbles are visible or if the liquid has overflowed over the edges and into the grooves, the chamber must be cleaned and feeding must be started again.

Blood diluting pipettes to be used:

• Leucocytepipette,shownabove(whitebulb)

• Erythrocytepipette,shownabove(redbulb)

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COUNTING THE PARTICLES

Counting Technique

Under the microscope, the lines of the counting grid appear dark.

Counting assumes precise knowledge of the limit lines of the counting chambers used. These are shown in the illustration.

To ensure cells which are on or along the limit lines are not counted twice or are not missed during the count, start the count at the top left corner and follow the direction shown by the arrow, as shown in above right illustration.

Notes on Countinga) The trim of the capacitor on the microscope must be almost closed for all chamber counts.b) The difference of the counter cells in the large squares and the group squares must not

exceed 10 cells.c) Double checks must be performed for all cell counts. After counting the two counting nets

the bottom counting net is to be counted in the same way as a check. When doing this it is to be ensured that the chamber has not dried out. This can be prevented by filling the bottom chamber only shortly before the count and the counting after the sedimentation time.

d) The difference between the totals of the counts for the two counting nets must not exceed 10 cells. The average value of the counts is then used in the calculation formula or multiplied by the corresponding factor.

CALCULATION

Number of cells per cubic millimeter

= Number of cells counted per square millimeter x dilution (if used) x 10

Number of cells per milliliter

= Number of cells counted per square millimeter x dilution (if used) x 10,000

One (1) Milliliter = 1000 cubic millimeters (cu mm)

One (1) Microliter (ul) = One (1) cubic millimeter (cu mm)

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CLEANING AND STORAGE

Cleaning

Before use and after completing each count, remove the cover glass and clean the counting chamber with water or a mild cleaning solution (10% solution of bleach). Dry the counting chamber with a soft cloth or wipe, or rinse with acetone.

Storage

These products should be stored in the original packaging, in a dry environment, at a constant room temperature, preferably protected from any sources of heat and light. Should a change in temperature be necessary, this should take place slowly. Take note of the applicable maximum stacking heights. Avoid strong jolting actions, also when transporting internally.

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www.grahamfield.com

GF Health Products, Inc. is not responsible for typographical errors. All illustrations, specifications, packaging and warranties contained in this literature are based on the latest product information available at the time of printing. The most current product information, including the most current version of these instructions, can be found online at www.grahamfield.com. Graham-Field and Grafco are registered trademarks of GF Health Products, Inc.

©2015GFHealthProducts,Inc.,2935NortheastParkway,Atlanta,GA30360,tel770-368-4700,fax770-368-2386•MadeinGermany

WARRANTYSCOPE OF WARRANTYGF Health Products, Inc. (“GF”) warrants to the original purchaser only that it will replace or repair components, at GF’s sole discretion, that are defective in material or workmanship under normal use and service. All warranties are conditioned upon the proper use of the products strictly in accordance with good commercial practice and applicable GF instructions and manuals, including proper use and maintenance. To the extent that a component is warranted by a third party, GF conveys all of its rights under that warranty to the original purchaser, to the extent permitted. This limited warranty shall only apply to defects that are reported to GF’s customer service team within the applicable warranty period and which, upon examination by GF or its authorized representative, prove to be a warranty item. This limited warranty is not transferable. Within the guidelines set forth in this document, this product is warranted for one (1) year. The applicable warranty period shall commence from date of shipment to the original customer, unless there is an expiration date on the component in which case the warranty shall expire on the earlier of warranty period or the expiration date.OBTAINING WARRANTY SERVICEThis limited warranty shall only apply to defects that are reported to the Distributor from whom the Customer purchased the product within the applicable warranty period. If there is not a Distributor, you must contact GF directly by calling 770-368-4700, sending a fax request to 770-368-2386, or by e-mailing a request to cs@grahamfield.com. Specific directions will be provided by the Customer Service Representative. Failure to abide by the specific directions will result in denial of the warranty claim.EXCLUSIONSThe warranty does not cover and GF shall not be liable for the following: 1) Defects, damage, or other conditions caused, in whole or in part, by misuse, abuse, negligence, alteration, accident, freight

damage, tampering or failure to seek and obtain repair or replacement in a timely manner;2) Products which are not installed, used, or properly cleaned and maintained as required in the official manual for the

applicable product;3) Products considered to be of a non-durable nature including, but not limited to: casters, filters, fuses, gaskets, lubricants,

and charts;4) Accessories or parts not provided by GF;5) Charges by anyone for adjustments, repairs, replacement parts, installation or other work performed upon or in connection

with such products which are not expressly authorized in writing, in advance, by GF;6) Any labor or shipping charges incurred in the replacement part installation or repair;7) Costs and expenses of regular maintenance and cleaning; and8) Representations and warranties made by any person or entity other than GF.ENTIRE WARRANTY, EXCLUSIVE REMEDY AND CONSEQUENTIAL DAMAGES DISCLAIMERTHIS WARRANTY IS GF’S ONLY WARRANTY AND IS IN LIEU OF ALL OTHER WARRANTIES, EXPRESS OR IMPLIED. GF MAKES NO IMPLIED WARRANTIES OF ANY KIND INCLUDING ANY IMPLIED WARRANTIES OF MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE. IF ANY MODEL OR SAMPLE WAS SHOWN TO THE CUSTOMER, SUCH MODEL OR SAMPLE WAS USED MERELY TO ILLUSTRATE THE GENERAL TYPE AND QUALITY OF THE PRODUCT AND NOT TO REPRESENT THAT THE PRODUCT WOULD NECESSARILY CONFORM TO THE MODEL OR SAMPLE IN ALL RESPECTS. THIS WARRANTY IS LIMITED TO THE REPAIR OR REPLACEMENT OF THE DEFECTIVE PARTS. GF SHALL NOT BE LIABLE FOR AND HEREBY DISCLAIMS ANY DIRECT, SPECIAL, INDIRECT, INCIDENTAL, EXEMPLARY OR CONSEQUENTIAL DAMAGES, INCLUDING, BUT NOT LIMITED TO: DAMAGES FOR LOSS OF PROFITS OR INCOME, LOSS OF USE, DOWNTIME, COVER, OR EMPLOYEE OR INDEPENDENT CONTRACTOR WAGES, PAYMENTS AND BENEFITS.The warranties contained herein contain all the representations and warranties with respect to the subject matter of this document, and supersede all prior negotiations, agreements and understandings with respect thereto. The recipient of this document hereby acknowledges and represents that it has not relied on any representation, assertion, guarantee, warranty, collateral contract or other assurance, except those set out in this document.For additional information on this product or this warranty, please contact a GF Customer Service Representative.NOTES:1) Additional terms and conditions may apply.2) Freight claims must be notated on the appropriate shipping documents and must be made with immediacy. International,

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3) Claims for any short shipment must be made within three (3) days of the invoice date.