MICROFIL Injection Compounds

Transcription

1 MICROFIL Injection Compound

2 MICROFIL compound will fill and opacify microvacular and other pace of non-urviving animal and potmortem tiue under phyiological injection preure. The continuou, cloed vacular ytem tend itelf to flow through injection or perfuion technique. Following injection, MICROFIL compound cure to form a threedimenional cat of the vaculature. MICROFIL MV-erie compound are available in five radiopaque color, a well a clear. MV-erie compound require either an alcohol-methyl alicylate or glycerin clearing equence, whereby the refractive index of the clearing olution i the ame a the refractive index of the tiue. Thi allow for microcopic examination of a elected vacular bed. MICROFIL CP-101 compound i intended for ue in cat-corroion technique, and i deigned for filling large blood veel (greater than 100 micron). Although the CP-101 compound will fill capillarie, thee veel fragment when the upporting tiue i removed through expoure to a potaium hydroxide olution. When cured, MICROFIL CP- 101 i milky white in color. Cat made uing CP- 101 will maintain their dimenional accuracy indefinitely. Advantage offered with MICROFIL compound over previouly available rubber injection material include: Complete filling with minimal hrinkage, to enhance veel continuity and to produce in cleared preparation a vivid, optically cleared pecimen that allow a precie tudy of the microcirculation. Color diverity, to provide delineation within the circulatory tree for microcopic examination and photographic illutration. Area of invetigation In phyiology, MICROFIL viualization provide a mean for etablihing the precie vacular architecture of pecific organ, allowing comparion between normal and abnormal tructure. In urgery, viualization of the microcirculation and microanatomy i leading to improved urgical technique in the repair of nerve, tendon, and blood veel. In gatrointetinal reearch, MICROFIL compound characterize and decribe change in vacular pattern aociated with everal pathological condition. Injected pecimen, when preerved in methyl alicylate or glycerin, alo erve a a definitive teaching adjunct. MV-erie mixing procedure To achieve a vicoity level uitable for injection of the microcirculation, it i neceary to blend the MV compound with an equal quantity (by weight) of MV-Diluent. Volume mixing require 5ml of diluent for every 4ml of compound. The mixture of compound and diluent i catalyzed with 5% (by weight or volume) of MV Curing Agent; Vicoity range from 20 to 30 centipoie. Working time i 20 minute and begin with the addition of curing agent. Table 1. Phyical propertie of MICROFIL compound MV- MV-112 MV-117 MV-120 MV-122 NW-130 MV-132 Diluent CP-101 Color White Orange Blue Yellow Red Clear Clear Milky White Specific gravity Vicoity, 1 centipoie Gel time, 2 minute Ueful Shelf life, Indefinite 6 month Note 1. Vicoity meaured with a Brookfield Model LVF Vicometer, with a No. 2 pindle at 30 RPM. 2. Gel time meaured on a blend (by weight) of one part MV compound and one part MV-Diluent followed by addition of 5% MV Curing Agent. Get time i time required for mixture to ceae flowing. 3. Shelf life i in exce of four month. For material held beyond four month, it i poible to run the following tatic tet to determine acceptability: a. Mix 5 gram of MV compound with 5 gram of MV-Diluent in a vial. b. Add 10 drop (medicine dropper) of MV Curing Agent. c. Cap, hake, and refrigerate overnight. d. The mixture hould gel in the vial after thi procedure to aure a cure in ubequent animal injection.

3 Catalyzed mixture will form an elatomeric gel after 90 minute at room temperature. Curing take place with non-exothermic cro-linking and minimal volume change. It ha been poible to refrigerate pecimen immediately after injection and till obtain complete cure after overnight aging. Thi procedure decreae odor level for ubequent ectioning. Perfuion technique Two technique of tiue clearing are decribed below. Alcohol-methyl alicylate clearing produce a tiffer tiue which, from an aethetic point, provide a pleaing view for gro obervation. Glycerin clearing produce a more flexible tiue, allowing eaier manipulation for-a given veel. Alcohol-methyl alicylate clearing Non-wetting feature of MICROFIL compound prevent any interaction with blood. Therefore, in the non-urviving animal, a elected vacular bed can be readily perfued without prior wahout of blood. Heparinization to maintain blood fluidity, however, ha been ued to realize improved injection preparation. For the injection of blood veel from vacular bed removed potmortem, wahout of clotted blood with aline i adviable. Selected vacular bed are perfued through their acceible artery and drained through a imilar vein. Infuion preure will vary with the animal mean ytemic preure. For organ from the dog, cat, rat, MICROFIL MV-130 (Red) injection of rat trachea. Courtey Robert A. Acland, Univerity of Louiville MICROFIL MV-130 (Red) injection of rat kidney. Courtey Robert A. Acland, Univerity of Louiville

4 Table 2 Alcohol-methyl alicylate clearing equence Firt Day Immere in a 25% olution of ethyl alcohol. Second Day Immere in a freh olution of 50% ethyl alcohol. Third Day Immere in a freh olution of 75% ethyl alcohol. Fourth Day Immere in a freh olution of 95% ethyl alcohol. Fifth Day Immere in abolute ethyl alcohol. Sixth Day Immere for 12 to 24 hour in methyl alicylate. If tiue ha not cleared, return to 95% ethyl alcohol tage and repeat final tep of clearing procedure. Note 1. At the 50% ethyl alcohol concentration, tiue pecimen may be bleached with 6% hydrogen peroxide for one day. After bleaching, continue the normal clearing procedure. Peroxide bleaching permit greater depth perception; however, thi procedure mut be conidered againt ome lo in color contrat. 2. In the final tage of ethyl alcohol-methyl alicylate clearing, the clearing liquid may have a cloudy appearance. Addition of a mall amount of 10% ethyl alcohol will alleviate thi condition. Cro-ectional view of monkey jejunum perfued with MICROFIL MV-118 (Maroon)* followed by MV-122 (Yellow). Courtey D. G. Reynold, Walter Reed Army Intitute of Reearch and man, a preure of 150mm. Hg for arterial filling ha been ued, and 25 to 50mm.Hg for venou filling. After the vacular bed i perfued and the MICROFIL injection ma allowed to cure overnight at room temperature, the tiue i ubjected to the clearing equence decribed in Table 2. Thin tiue may be cleared without ectioning, but thicker organ, uch a kidney or brain, hould be cut into 1-centimeter lice before immerion. The alcoholmethyl alicylate clearing technique i applicable to all type of tiue with the exception of brain tiue. In the cae of brain tiue, it i neceary to allow two day for each tep, with an alcohol olution change every day. Glycerine clearing The animal i anethetized with Nembutal, 25 mg/kg i.v., at the ame time it i heparinized, to enure effective removal of it blood volume during perfuion. A midline inciion expoe the abdominal vicera from the ternal notch to ymphyi. Following the placement of an abdominal retractor, the thoracic cage i opened rapidly, the thoracic aorta iolated, and a polyethylene cannula inerted ditally. The arterial cannula i connected to a ine wave perfuion pump and, prior to intigating perfuion, the right atrium i opened to erve a a drain vent. The animal i perfued with aline until all of the viceral blood volume i fluhed out and the perfuate drained through the arterial vent i eentially free of blood. Adequate perfuion i characterized by evere blanching of all viceral organ. During perfuion, the curing agent i added to the MICROFIL injection ma. When perfuion i complete, the ilicone rubber (e.g., MV-130 Red) i infued through the aortic cannula by yringe. When filling i complete, all organ have a rich, red coloration. MICROFIL compound infuion i continued until the injection ma flow freely from the atrial vent. The atrium and arterial cannula are then clamped and the animal i placed under refrigeration at 4 C overnight, to allow polymerization. On the following day, pecimen are taken by careful diection, and placed in a 50% mixture of water and glycerin. At ucceive 24-hour interval, the glycerin concentration i raied to 75%, then 85%, and finally pure glycerin. Thi procedure clear the tiue o that microcopic examination readily allow three-dimenional viualization of the vacular bed. *MV-118 (Maroon) i no longer available and ha been replaced with MV-130 (Red).

5 Additional Note 1. All MV-erie compound are compatible with one another. Therefore, it i poible to mix color together to uit your need (e.g., mix MV-120 Blue with MV-122 Yellow to produce a green compound). 2. Fater cure rate are poible by replacing the conventional MV Curing Agent with 2% ethyl ilicate and 1% tannou octoate. Uing thi crolink and curing agent combination decreae working time to 5 minute with complete cure in 20 minute. If you deire thi type of cure ytem, pleae pecify when placing your order. There i no extra charge for ubtituting thi cure ytem in place of MV Curing Agent. 3. Occaionally it may be neceary to decreae vicoity by changing the mix ratio to either 2 or 3 part MV-Diluent for each part MV compound. If your tudy require uch action, the correct level of MV Curing Agent i 10% (by weight) of the amount of MV compound ued. 4. One procedure for veel differentiation i to completely fill a given circulation with conventional MV compound (e.g., MV- 130 Red) then, once the circulation i filled, immediately re-inject the artery with a high-vicoity verion of MV compound in a different color (e.g., MV-120 Blue). Thi i accomplihed by ubtituting MV-Diluent, which ha a vicoity of 5 cp, with HV-Diluent, which ha a vicoity of 1000 cp. With HV-Diluent, the vicoity of MV-130 Red increaed to cp. Although thi procedure topped penetration at the capillary level, complete ucce wa obcured by hunting activity. If you deire to replace MV-Diluent with HV-Diluent, pleae pecify when placing your order. There i no extra charge for thi ubtitution. 5. Store MICROFIL kit at room temperature for maximum retention of pigment diperion. Keep all container tightly capped. If pigment ettling occur, it i better to lightly hake the MICROFIL compound container and decant thi portion. Stirring the container may put an agglomerated pigment particle into the ytem, which can be detrimental to perfuion. 6. If catalyzed material hould pill on clothing, the bet available olvent i MV-Diluent. To facilitate removal of cured compound, welling and oftening will occur on contact with an aromatic olvent uch a toluene or xylene, and chlorinated olvent uch a trichloroethylene. Arterial injection of a dog kidney uing MICROFIL MV-112 (White). Demontrate filling of glomeruli and peritubular capillarie at 50x magnification. Courtey A. C. Berger, Harvard Medical School Vaa vaorum of the human coronary artery filled with MICROFIL compound. Courtey of A. C. Barger, R. Beeuwke 111, L. L. Lainey and K. J. Silverman, Harvard Medical School

6 Cover Photo: MICROFIL injection of the left coronary artery of the human heart. Courtey of A.C. Barger, R. Beeuwke III, L.L. Lainey and K.J. Silverman, Harvard Medical School Ordering Information When ordering material, pleae remember kit weight i baed upon the combined weight of MV compound and MV-Diluent (i.e., a 1-pound kit contain 8 ounce of MV compound and 8 ounce of MV-Diluent). In addition, each kit contain enough MV Curing Agent to cure the content. Kit Size Specification 1 lb. Any one color 2 lb. Any one or two color 8 lb. (1 gallon) Any combination of color MICROFIL injection kit are available from: Flow Tech, Inc. P.O. Box 834 Carver, Maachuett Tel: (508) Fax: (508) Term: Net 30 FOB Carver A bibliography i available on requet. While the information herein i believed to be reliable, Flow Tech doe not guarantee it accuracy. Uer are urged to perform their own tet. MICROFIL product are old without warranty, and variou patent may be pertinent to their ue and to the ue of compoition containing them. The information contained herein i not intended a a recommendation to ue MICROFIL product o a to infringe on any patent. Flow Tech aume no liability for the uer violation of patent or other right. 1996, Flow Tech, Inc. All right reerved. MICROFIL i a regitered trademark of Flow Tech, Inc., Carver, Maachuett.