The present status of sale

• Stable liquid and water-absorbable sponge made by KytoGEL is available.
• You can voluntarily select water, saline, PBS, and medium as solvents.

喝喝KytoGEL liquid may be insoluble in pH 6 or more.
喝喝The opportunity of insolubilizing KytoGEL increases at pH 7.0 or more.
喝喝Please adjust the concentration by using 4 % of aqueous KytoGEL when using
BIBBiIBspecial medium or buffer.

• When you adopt a special medium or buffer, you should dilute 4 wt% aqueous KytoGEL with the medium or buffer of 2-fold concentration.
• Please prepare commercially available device for UV irradiation.IUV irradiation

• Please consult us for ordering articles specially made such as KytoGEL sponge including antibacterial and antiviral agent. Inquiry form

• Lectin method to precisely isolate erythroblasts consists of the following process.
• Single density centrifugation: removal of erythrocytes and a part of granulocytes (about 40 min.)
• CD45 depletion: removal of leukocytes (about 40 min.)
• Lectin method: Isolation and enrichment of erythroblasts (about 50 min.)
• Cytospin: fixation and drying of cells on the slide glass (about 5 min.)
• May-Grünwald-Giemsa (MGG) staining: morphological staining of cells on the slide glass (10 min.)
• Operation of ISOLECT is carried out at ordinary room temperature (18~22℃).

• ISOLECT will supply more FISH-reactive erythroblasts than other isolation technologies such as MACS, filtration, or microchannel.
• We have so far conducted erythroblast purification using ordinary centrifugation and leukocyte depletion with magnetic beads. In the course of the optimization concerning their conditions, enucleation of erythroblasts caused by excessive centrifugation and stirring with stiff microspheres had been frequently observed.
• Several researchers reported that erythroblasts which were isolated from maternal circulation are FISH-impervious. On that occasion, the magnetic beads immobilizing anti-CD45 and/or CD71 antibodies had been adopted as isolation method.
• ISOLECT does not contain the process of magnetic beads.

Lectin method

Equipments which are provided by customers to carry out ISOLECT

• Low speed centrifuge with swing bucket rotor (required)
• Cell counter
• Incubator (18℃) or experimental room (18 to 22℃)
• Microscope (required)
• Image analyzer, laser micro dissection

• Image analyzer Metafer4 (MetaSystems) is attached to AXIO II (Carl Zeiss). Metafer manages auto-stage of AXIO and indicates morphological erythroblast candidates which are subject to MGG staining. The position of cells which were selected as erythroblast are computationally memorized.
• Laser captured microdissection PALM (Zeiss) holds the position data of Metafer in common, and then easily catapults and collects erythroblasts as a single cell.
• DNA arising from recovered erythroblasts are amplified, and its mutation or lack is subsequently analyzed.
• Metafer and PALM enabled to simultaneously detect Y FISH signals and amplified SRY (Sex-determining region Y) from erythroblasts of same pregnant woman carrying male fetus.
• PCR revealed paternal DNA from the lectin-collected fractions including all cells which adhered to the glass slide. This fact indicates that in order to detect defects or mutations in DNA, single-cell manipulation is not essential.
• When applicability of digital PCR to quantitate absolutely the copy number of target from a small quantity of DNA arising from rare cells has been proved, not only prenatal test of congenital anomalies but also fetal therapy will expand in the future.

• Chamber slides

• A chamber having two wells is mounted on a glass slide.
• Two-well chamber is applicable to the assay to simultaneously use different type of FISH probes.
• Size of the well: 22.5 x 22.6 mm.
• EOG-sterilized

• Cytospin device

Inquiry form

• G-coat ware is a dish, chamber slide, or microsphere cated with neoglyconoid G-CPS composed of oligosaccharide-carrying polystyrene.　Neoglyconoid
• Soap-free latex is made by polystyrene, and their particle size has been kept uniform.
• Table described below indicates G-CPS which is currently available.
• Please inquire a specific modification involving the plural G-CPS coating or the combination coating of HCPS and collagen, etc..

• G-CPS list

• Polystyrene dishes
• IIII－EOG-sterilized
• Chamber slide
• IIII－Smooth and flat surface glass slide (non-fluorescent), EOG-sterilized
• Soap-free latex
• IIII－Latex modified with dye or fluorescence is also available.

Inquiry form