An Automatic Multiple Diluting Machine

S, Baron; Bl, Burch; Bw, Uhlendorf

doi:10.1093/ajcp/36.6_ts.555

American Journal of Clinical Pathology

1961

DOI: 10.1093/ajcp/36.6_ts.555

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An Automatic Multiple Diluting Machine

Baron S

Burch Bl

Uhlendorf Bw

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1972

1981

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“…Assay of each sample of interferon requires the use of as few as 6 tube cultures-i.e., 3 dilutions of interferon, using 2 tubes/dilution. The hemagglutination assay is inherently fast and simple (1 1) and can be easily automated ( 12). Finally, this interferon assay is highly sensitive as evidenced by the high titers of the reference interferons.…”

mentioning

confidence: 99%

Improved Assays for a Variety of Interferons

Oie¹,

Buckler²,

Uhlendorf³

et al. 1972

Experimental Biology and Medicine

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Interferon assays are based on the inhibition of either viral function or synthesis of virus or its components. No one assay has proven to be ideal but significant improvements are constantly being introduced ( 1 ) . A study was initiated to develop a widely applicable assay which would combine rapidity, simplicity and reliability and which is based on the inhibition of virus multiplication during a single cycle of growth.Materials and Methods. Cell cdtures. Mouse L cells (obtained from Dr. J. S. Youngner, University of Pittsburgh) primary and secondary mouse embryo cells (NIH Swiss), primary chicken embryo cells and 2 diploid human cell lines (MA-308 and RCTC-2) were cultured to confluency by standard procedures. For assay, cells were cultured in 16 x 150 mm tubes, held vertically in a spring rack ( 2 ) and covered loosely with a single metal lid (3 ) .Viruses. GD-7 virus, a mouse picornavirus, was produced and plaque-titrated [ 106.9 plaque-forming units (pfu)/ml] in BHK-2 1 cells as previously described (4). Sindbis virus was prepared and plaqued pfu/ml) in chicken embryo monolayer cultures. Vesicular stomatitis virus (VSV) , Indiana strain, was also prepared in chicken embryo cells but the plaque titration was done in mouse L cells ( 107v5 pfu/ml).Interferons. Mouse serum interferon (MSI) ( 5 ) titered 104s2 units/ml in mouse L cells. Human interferon was prepared from human diploid fibroblast cell cultures infected with Chikungunya virus [ input multiplicity of infection (MOI) = 11 and it titered 103.0 units/ml in MA-308 cells. Chicken interferon induced in ovo with NWS influenza virus (6) titered 102a3 units/ml in chicken embryo cells. These titers were obtained using the methods described below.Hemagglutination assay. Titration of GD-7 virus hemagglutinin (HA) was done by standard methods as previously reported (4). 0.06% fetal bovine serum (FBS) was added both to erythrocyte suspensions and to the virus diluent because it was found to be essential for the proper development of the hemagglutination patterns. Titration of Sindbis virus HA was done as previously reported (7) using the determined optimum pH of 6.0 and a 0.125% concentration of gander erythrocytes in the final reaction mixture. Results. Time course of H A production.To determine the times of maximal HA production in the virus-cell systems under study, single step growth curves were determined (Fig. 1 ) . It may be seen that maximum HA production occurred at 1 2 hr for Sindbis virus in chicken and mouse embryo cells, at 16 hr for Sindbis virus in human MA-308 cells and 18 hr for GD-7 virus in mouse L cells.The time course of viral production in interferon treated cells is generally not different or only slightly delayed (8). Therefore, during interferon assays lfluids were harvested for HA determinations 18-24 hr after infection.Interferon assay b y inhibition of yield of viral H A . Duplicate or triplicate tube cultures of cells were each exposed to l ml of interferon diluted in Eagle's minimal essential medium (EMEM) plus 2% FBS. After 18-24 ...

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mentioning

confidence: 99%

Improved Assays for a Variety of Interferons

Oie¹,

Buckler²,

Uhlendorf³

et al. 1972

Experimental Biology and Medicine

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Das Autoselect‐System – ein Automatensystem zur Selektion von Antibiotikaproduzenten. IV. Achtfach‐Verdünnungsautomat

Schicht¹,

Peissker²,

Steininger³

et al. 1981

Acta Biotechnologica

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As a part of the Autoselect-system an eightfold diluter is described. Cassettes with 64 tubes are placed on the carriage of the machine and moved automatically in eight steps. One of the cassettes is loaded with separated samples in 64 tubes and the other one with 64 empty tubes. When the carriage stops, a defined volume of samples is exhausted by eight pipettes, mounted on a bridge spanning over the ground unit from left to right, and after beeing moved to the second cassette the pipettes deliver the samples into a row of empty tubes. At the same time by eight syringes mounted on both sides of the machine a defined volume of buffer is delivered through tubes and canules. By this way all samples can be diluted in two minutes. ZnsammenfassungEin Achtfach-Verdunnungsautomat a1s Teil des Autoselect-Systems wird beschrieben. Auf einem Transportwagen wird eine Kassette mit 64 Proben und eine weitere mit 64 leeren Rohrchen in 8 Arbeitsschritten bewegt. Der Verdiinnungsvorgang 18;uft folgendermaBen ab : -Entnahme eines definierten Volumens der Antibiotika-Kulturlosungen aus einer Reihe von Proben mittels 8 Pipetten, die von einem Pipettenwagen Lhnlich einer Briicke quer iiber die Grundeinheit angeordnet sind. -Nach Transport des Pipettenwagens wird die Fliissigkeit in leere Rohrchen der anderen Kassette entleert; gleichzeitig wird aus 8 Spritzen, die seitlich angeordnet sind, eine definierte Menge Puffer zur Verdiinnung in die R6hrchen dosiert. -I n angegebener Weise werden alle 64 Proben innerhalb von 2 Minuten verdiinnt.

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Das Autoselect‐System – ein Automatensystem zur Selektion von Antibiotikaproduzenten. V. Achtfach‐Pipettierautomat

Steininger¹,

Schicht²,

Peissker³

et al. 1981

Acta Biotechnologica

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SuniniaryAr, automatic eightfold-pipetter has been developed with the same basic unit as the diluter of the Autoselect-system. The pipetter is fitted with eight pipettes on a bridge over the basic unit. By means of the pipettes it volume of 0.05 ml from eight tubes is exhausted and delivered in the holes of a test plate. Both the test plate and t i cassette with 64 tubes are located on a moving carriage. the test plate on the first floor and the cassette on the ground floor. If the pipettes transfer the stmplss from the tubes to the holes the distance between the pipettes muat be changed by . G speci.11 device from 20 to 30 mm, because the distance between the holes on the test plate differs from that of the tubes. For the transfer of 64 samples 2 minutes only are needed. ZusarnrnenfassungEin Achtfach-Pipettierautomat mit der gleichen Grundeinheit wie der des Verdunnungsautomaten des Autoselect-Systems wurde entwickelt. Y Pipetten, die auf einer Brucke nngeordnet sind, entnehmen den Probenrohrchen jeweils 0,05 ml Kulturflussigkeit und geben sie in die Locher der Testplatte ab. Die Testplatte und die Kassette mit 64 Proben sind auf einem Transportwagen verankert. Durch eine spezielle Vorrichtung wird der Abstand zwischen den Pipetten wiihrend des Transports vergroBert, urn den Abstiinden der vorgestanzten Locher der Testplatte zu entsprechen. Fur die ttbertragung von 64 Proben werdeii 2 Minuten benotigt.

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An Automatic Multiple Diluting Machine

Cited by 3 publications

References 0 publications

Improved Assays for a Variety of Interferons

Improved Assays for a Variety of Interferons

Das Autoselect‐System – ein Automatensystem zur Selektion von Antibiotikaproduzenten. IV. Achtfach‐Verdünnungsautomat

Das Autoselect‐System – ein Automatensystem zur Selektion von Antibiotikaproduzenten. V. Achtfach‐Pipettierautomat

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