Pathology Research 2Ol 3 D-1. Lemosol'and Lemosol@ A (Ace) ................9 G-1. General Staining ....'...'............14 G-2. Polysaccharide Staining ..............................'16 G-3. Connective Tissue Staining ..'...'......'....'....19 G-4. Staining of Fibrins and GlialTissues.. .. """""""."..'."23 G-5.IntracellularInorganicSubstanceStaining .".'..'.'..23 G-6. Cytological Staining ....... ... ... """ ""'. "'24 G-7. Blood Cell Staining ... .. ... .25 G-8. Hard Tissue Staining (TRAP/ALP Stain Kit) .....................26 G-9. Other stains .'.'."'...'.'..'.............27 Itir K-1 H stoMap Ser es g2 A cridine Oranqe 7 ne Yellow Alcian Blue Solution. oH 2.5 ne Blue . Oranqe G Solution ne Blue Solution 27 ne Blue-Water Soluble 28 v's Mountinq Media, Soluble Dootosis m situ Detection Kit tvako 29 oootosis Ladder Detection KiIWako Auramine Azocarmine G Solution B Berlin Blue Stainino Set Bismarck Brown 5 Bouin Solution 28 c Canada Balsam 7 Carmine 5 Carnov Solution 15 Caft azzi's Hematoxvlin Solution 15 Carrazzi's Hematoxvlin Solution (x2) 27 Chlorazol Black E 6 Cold Schiff's Reaoent Collodion (5%) Collodion (10%) I Colloidal lron Stock Solution 27 Conqo Red 27 Crvstal Violet (DAB D DAB Tablet . 4HCl Smo/Tablet) 29 DAB Tablet (DAB . 4HCl 1Omo/Tablet) 29 DAB TRIS Tablet. oH7.6 Decalcifvino Solution A (Plank-Rvchlo orotocol) 8 Decalcifvinq Solution B (EDTA Drotocol) 27 4'.6-Diamidino-2-ohenvlindole Dihvdrochloride n-Hvdrate 30 DNA lsolator PS Kit 30 DNA lsolator PS-Raoid Fleaqent 1t Dodecenvlsr rccinic Anhvclricle tDDSAI 15 E Eosin Alcohol Solution. acid extract 27 Eosin Y 5 0.1% Eosin Y Ethanol Solution 2% Osmium fflID Oxide Solution 5 0.5% Eosin Y Ethanol Solution 4% Osmium ffllj) Oxide Solution 1% Eosin Y Solution Eoon 174 Eoon 81 2 Epon 81 5 27 Ervthrosine 30 Ethanol 5 Buffer Solution 11 Ether 11 546 27 Fast Blue RR Salt F 11 568 10% Formalin Neutral Buffer Solution 580 3 15% Formalin Neutral Buffer Solution 2O%o Formalin Neutral Buffer Solution d 5% Formic Acid (for Decalcificatior 27 Fuchsin Acid 27 Fuchsine Basic 11 G Gelatin 11 Gelatin. Caosules No.0 11 Gelatin. Caosules No.00 27 Gentian Violet R 25 Giemsa Stain Solution 5% Sodium Sulfate Solution 6 1 0% Glutaraldehvde Solution 6 20% Glutaraldehvde Solution 6 70% Glutaraldehvde Solution H Hematoxylin Monohydrate 32 HistoMap Mouse Normal Cerebellum, Paraffin Embedded Tissue JZ HistoMap Mouse Normal Cerebrum, Paraffin Embedded Tissue Section HistoMao Mouse Normal Heart. Paraffin mbedded Tissue Section oz HistoMao Mouse Normal Kidnev Paraffin ed Tissue Section HistoMao Mouse Normal Liver. Paraffin nbedded Tissue Section HistoMao Mouse Normal Luno. Paraffin nbedded Tissue Section HistoMap Mouse Normal Pancreas, Paraffin Embedded Tissue Section 0.05% Toluidine Blue Solution 32 HistoMao Mouse Normal Rectum, Paraffin Embedded Tissue Section 0.05% Toluidine Blue Solution JZ HistoMao Mouse Normal Soleen Pare mbedded Tissue Section 0.05% Toluidine Blue Solution JZ HistoMap Mouse Normal Stomach, Paraffin Embedded Tissue Section HistoMao Mouse Normal Testis Paraffin nbedded Tissue Section 5% Trichloroacetic Acid Solution (for Decalcifica JZ HistoMao Rat Normal Cerebellum Para mbedded Tissue HistoMao Rat Normal Cerebrum Pare rmbedded Tissue Section HistoMap Bat Normal Head, Paraffin Embedded Tissue Section Van Gieson Solution F 32 HistoMap Ra't Normal Kidney, Paraffin Embedded Tissue Section JZ HistoMap Rat Normal Liver, Paraffin Embedded Tissue Section 32 HistoMap Rat Normal Lunq, Paraffin Embedded Tissue Section JZ HistoMap Bat Normal Pancreas, Paraffin Embedded Tissue Section 32 HistoMao Rat Normal Rectum. Paraffin Embedded Tissue Section 32 HistoMao Rat Normal Soleen aratfin Embedded Tissue Section 32 HistoMao Rat Normal Stomach. Paraffin :mbedded Tissue Section 32 HistoMap Rat Normal Testis, Paraffin Embedded Tissue Section 8 1O% Hydrochloric Acid (for Decalcification) f ixation means a process by which when observing microscopically the ecological or pathological state of a tissue I specimen taken from a dead or live body, the tissue specimen is preserved in as "intact" a state as possible. The objectives are to: Stabilize the main constituent protein of the tissue specimen as fast as possible to stop autolysis Minimize the degeneration and deformation of the tissue specimen caused by the effects of chemicals and heat during the subsequent specimen preparation process. for Light Microscopy Neutral buffered formalin Formalin solution is used in large quantities as a routine fixative in the preparation of tissue specimens for light microscopic observation. Formalin is decomposed to produce formic acid, which may affect some tissues. This neutral buffered formalin is adjusted to pH 7.4 approximately by the addition of sodium phosphate to formalin to eliminate the effect of acid on tissues. Not only is neutral buffered formalin a superior fixative for routine lab work, it is also beneficial for a number of special purposes. For example, neutral buffered formalin can be used to a certain extent for histochemical detection of enzymes, hemoglobin and other molecules; for fixing mucopolysaccharides, glycogens, neurofibrils and other tissue components that cannot be treated with regular formalin; and for electron mtcroscopy. Formaldehyde Product Name Composition pH Penetration Strength Content 10% Formalin Sodium phosphate, monobasic, Dihydrate ......4.5 g O Penetration strength is in 3.7-4.3 w/wo/o 7.4-7.5 Neutral Buffer Solution Disodium phosphate 6.5 g the order of 2O%o > 15%o > Prepare the 1 L solutjon to add ion-exchanged water 10%. O Specimens must be placed in the 10% fixative 15%o Formalin Sodium phosphate, monobasic, Dihydrate . 4.5 g 5.6-6.4 w/w%6 7.35-7.45 for several days or more. Neutral Buffer Solution Disodium phosphate . ..6.5 g OThe 20o/o fixative allows Prepare the 1 L solution to add ion-exchanged water fixation in one or two days, 20o/o Formalin Sodium phosphate, monobasic, Dihydrate......4.5 g halfway between the 10% 7.5-8.5 wlw%o 7.3-7.5 Neutral Buffer Solution Disodium phosphate 6.5 g and 2o%io solutions. Prepare the 1 L solution to add ion-exchanged watel Product Name Grade Package Size Wako Cat. No. 1L 062-01 661 10% Formalin Neutral Buffer Solution for Tissue Fixaiion 20L 060-01 667 1L 069-02391 15% Formalin Neutral Buffer Solution for Tissue Fixation 20L 067-02397 1L 060-01 721 2Oo/o Formalin Neutral Buffer Solution for Tissue Fixation 20L 068-01727 Mildform Mildform products, fixatives for pathological tissues, are neutral buffered formalin solution prepared according to Lillie's formulation to which wine extract* is added. The wine extract minimizes the irritating and unpleasant odor of formalin, however, with regard to operational safety, minimum degree of odor is present so that formalin can be recognized. * Mechanism of wine extract's effect: The masking effect minimizes the irritating and unpleasant odor of formalin Features .-r Most suitable for tissue fixation of immunohistochemical staining by an immunoenzymatic technique. '' Mildform can preserve tissue in the advanced stages of autolysis without compromising visual clarity. ' Stable for more than 2 years at room temperature r' This solution, which lacks the unpleasant odor of conventional formalin, is useful for biopsies, experimental tissue sections and surgical resections. lt is also ideal for quick fixation of anatomical tissue in the operating room or pathology lab. ", The fixation and osmosis are equivalent to that of neutral buffered formalin solution or oreater. Pathology Research 201 3 A.-1. for Light {./lieraseopy Mildform Data 20% Formalin Neutral Mildform 20N Fixation Comparison penetration/fixation Buffer Soln. Fixation of rates for human liver tissue 1234 5678 10 12 14 16 18 20 22 24(time) 1. Mildform 1ON (Equivalent to 10o/o Formalin Neutral Buffer Sotn.\ 2. Mildform'10NM 3. Miidform 20N (Equivalent to 20o/o Formalin Neutrat Buffer Soln.) 4. Mildform 20NM 5. 10% Formalin (Source: Pathology Depaftment, Osaka Koseinenki n Hospital) Soln. 6.2O%o Formalin Neutral Buffer Soln. Product Name Grade Package Size Wako Cat, No. 1L 1 33-1 031 1 Mildform 10N for Pathology Research 20L 131-10317 1L 132-14301 Mildform 15N for Pathology Research 20L 1 30-1 4307 1L 1 36-1 0041 Mildform 20N for Pathology Research 20L 134-10047 | 0521 Mildform 10NM for Pathology Research E 20L 130- 10527 1 L 139-14311 Mildform 15NM for Pathology Research 20 L 137 4317 1L 1 39-1 0531 Mildform 20NM for Pathology Research 20L 137 o537 o N: signifying neutral. Mildforms are produced based on neutral buffered formalin. M' a Addition of methanol enhances osmosis into and fixation to tissues. Methanot is added to the product al 10% amount:91'l,v:l9,Ti"l1T^91^"_"1!:!1. of the package size O In general, NM-type is most suitable for rapid fixation with minimum length of time needed. Also suitable as a formatin-methanol fixative for fixing samples containing a large amount of connective tissues and adipose tissues. <Note> Although Mildform lacks the unpleasant, acerbic odor of formalin, it is still as toxic as regular formalin and therefore should be handled with oue care. Other Fixatives Product Name Composition Features This product allows fixation of small sections in 1 to 2 hours because of its strong penetration. Saturated Picric Acid Solution ,,,...... ..750 mL Fetal bone tissue can be thinly sliced because the solution's Bouin Solution Formalin Stock Solution .,........ ..250 mL decalcifying effect is relatively weak. This fixative is used Acetic Acid, Glacial ...... ... ,,SO mL frequently for endocrine tissue, glycogens and polysaccharides. After fixation, wasn specimen wilh 70% ethanol until the yellow color disappears. With better penetration pedormance than anhydrous alcohor, this product is suited for the fixation of glycogens and nucleic acids. lt can fix 3 to 4 mm specimens in a matter of 2Io 4 Carnoy Solution hours. After fixation, transfer the specimen to pure alcohol to remove chloroform and acetic and Acetic Acid, Glacial . .... ...... .100 mL acid, then embed, Because this product has strong dehydrating and delipidating proper- ties, overfixation will lead to extreme contractlon and hardening of the specimen.