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Cell Separation and PAP PEN
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NanoPhotometer™
Microparticles
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Cell Separation and PAP PEN

 
PAP PEN
PAP PEN is designed to provide a water repellant barrier when a circle is drawn around a specimen. The barrier of the PAP PEN retains antisera within the defined area and ensures, that only the amount of antobody needed for sufficient reaction is used. The PAP PEN stops spreading, reduces waste and divides slides into discret areas for immunostaining, histology, cytology, cell culture, virology, microbiology. The PAP PEN is effective for immunostaining, PAP method, ABC method, ASD method, enzyme and frozen section methods.
The staining is unsoluble in aceton and alcohol and can be removed if desired by xylene after the staining procedure.
Cat. #DescriptionEuroUnits
MKP-1Pap Pen Immunostaining Pen46,00
MKP-2Pap Pen Immunostaining Pen, extra thick59,90
 
Neubauer Improved / Disposable Hemocytometer
The Neubauer Improved - C-Chip is a disposable plastic one-time-use hemocytometer used for manual cell-counting. It consists of two surface patterned chambers with two parts for sample injection. The Hemotocytometer is made from plastic with glass-characteristics for standarized countings.
The handling is easy, 10 µl of cell suspension will be injected on the chip and counted. The grid pattern is exactly the same as the "Neubauer Improved" chamber. It consits of nine large squares of each 1 x 1 mm with 0,1 mm depth. Each square has a total volume of 0,3 cmm or 10E-4 ccm. The central square is divided into 25 small squares with triple lines and four corner squares are divided into 16 smaller squares

Methology
1. General methods with C-Chip: mix the samples well, load 10 µl of sample into the sample injection area so that the chamber is filled by capillary action, count the cells under the microscope

2. Mammalian-Cell Counting: treat the cell samples with trypsin-EDTA, carefully remove the supernatant with a pipett tip and avoid to disturb the pellet, add an appropriate volume of growth medium or PBS to dilute to a final concentration of 5 x 10E3 cells / ml to 5 x 10E6 cells/ ml

3. Erythrocyte counting ( 1:200 dilution): dilute blood using accepted laboratory methodes, load 10 µl deluted sample into the injection area, count the erythrocytes in the five small squares ( four small corner-squares and one small middle square) of the large center square

4. Leukocyte counting (1:20 dilution): dilute blood using accepted laboratory methodes, load 10 µl deluted sample into the injection area, count the leucocytes in the four large corner squares.
cat.#DescriptionPack sizeEuroUnits
M-NZC-Chip50 Chips78,00
 
Blood Separation Filter Tubes for Separation of Mononuclear Cells (MNC)
The 50 ml and 15 ml blood-separation-filter tubes are designed to separate mononuclear cells (MNC). This method has been developed by ESQUIRE(Leuco-sep).
  • Protocol to be performed under normal room temperature.
  • 1. Pipet 16 ml Lymphoprep* into the BLOOD-SEPARATION tube. (3ml for 15ml tube)
  • 2. Centrifuge at 1000 G for 30 seconds at room temp.
  • 3. Pour 15-30 ml freshly anticoagulated blood into the tube(3-8ml for 15ml tube). The blood does not have to be diluted, but can be diluted directly into the tube with a balanced salt solution.
  • 4. Centrifuge at 1000 G for 10 min. (or 800 G for 15 min.)
  • 5. After centrifugation the sequence of the tube content will be as follows:
    Plasma-MNC-Lymphoprep-Filter-Lymphoprep-Erythrocytes.
  • 6. MNC can be collected and transfered by a Pasteur pipet,or directly poured to another tube.
  • 7. Wash the MNC by adding 10 ml PBS and resuspend the cells by gentle aspiration with a pasteur pipet. Centrifuge at 250 G for 10 minutes at room temperature.
  • 8. Repeat step twice, resuspending the pellet in 5 ml PBS-buffer.
  • 9. The Lymphozites are now ready to pickup and stored in cryogenic vials.
    *Separation-solution is available as: Lymphoprep (Nycomed Amersham)
    Cat. #DescriptionPack unitEuroUnits
    M-BST1515 ml blood separation tubes, sterile10 x 10 tubes249,00
    M-BST5050 ml blood separation tubes, sterile4 x 25 tubes144.80
     
    Combed, Scrubbed Nylon Wool Fiber, Type R Separation of T-cell from B-cell lymphocytes with Nylon Wool Fiber
    Mononuclear leukocyte concentrates from freshly drawn blood can be separated into T-cell and B-cell enriched fractions using "Nylon Wool " Mononuclear lymphoid cells may be preseparated from the polymorphonuclear (PMN) leukocytes and contaminating erythrocytes by centrifugation on an isopynknotic solution of IsoPrep (specific gravity 1.077 gm/liter). If an isopynknotic solution is not used, the polymorphonuclear (PMN) leucocytes will adhere tightly to the nylon wool, while any erythrocytes will be eluted with the nonadherent cell fraction. Prior to the fractionation procedure, the mononuclear cells are washed once in Hank`s Balanced Salt Solution, counted and resuspended in Hank`s Balanced Salt Solution supplemented with 10% fetal calf serum (FCS) or in other cell growth media like RPMI 1640 with 10% FCS, Earl`s Saline with 10% FCS or Dulbecco`s PBS with 5% FCS.

    Our Nylon wool works well with e.g. human, mouse and rat cells. Please see also: current protocols in Immunology (1992) 3.2.1-3.2.4, contributed by K.S.Hathcock, T-Cell Enrichment by nonadherence to Nylon, John Wiley & Sons, Inc.
    Cell Separation Procedure

    1) T-cell enriched fraction
    Pack tightly 0,5gram of sterile Nylon wool to a 10 ml sterile syringe. Wash and equilibrate the Nylon Wool with selected media HBSS or RPMI 1640 with 10% FCS.
    1-2 x 10E8 mononuclear cells suspended in 2 ml of media are pipetted onto 0,5 grams of sterile Nylon wool.
    Incubate the syringe at 37°C for 45 to 60 minutes.
    Collect nonadherent T-cells by washing with HBSS and resuspend. Do not plunge.
    B-Cells are reduced in the syringe up to 3%, larger numbers of cells may be separated, when the amount of nylon wool (e.g. 1 g) and the size of the syringe is proportionally increased or by repeating the procedure.

    2) B-cell enriched fraction

    After elution of the not adherent T-cells from the Nylon Wool Fiber, the syringes are stoppered and filled with HBSS supplemented with 10% FCS. The syringes are opened and sterile plungers of the syringes are used to force the medium through the Nylon Wool Fibers. The cells are washed once in in HBSS and resuspended.
    Cat. #DescriptionPack sizeEuroUnits
    MKN-10Nylon Wool Fiber10g42,70
    MKN-50Nylon Wool Fiber50g136,00
    MKN-100Nylon Wool Fiber100g240,00
     
    Slide Boxes colored, for 100 Slides
    The slide boxes have cork liming on the bottom, to protect the slides. On the inside cover of the box is a corresponding inventory slide sheet. The boxes can be stored securely on top of each other and are closed with a rust resistant nickel plated clasp and hinge pin.Dimensions are 222 x 171 x 33mm ( LxWxH )

    Optional we offer stainless steel freezer racks to store the slide boxes in upright freezers for longtime storage of temperature sensitive samples. You find infos and prices in the chapter CRYO PRODUCTS.
    cat.#ColorPack sizeEuroUnits
    R100-Bblue14,95
    R100-Ggreen14,95
    R100-Yyellow14,95
    R100-Rred14,95
    R100-Wwhite14,95
    R100-GRAgrey14,95
    R100-SCHblack14,95
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    İ 1999 - 2011 Dipl. oec. troph. Gerlinde Kisker