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PubMed1997

Comparison of five incubation systems for rat liver slices using functional and viability parameters.

Precision-cut liver slices are presently used for various research objects, e.g. to study metabolism, transport, and toxicity of xenobiotics. Various incubation systems are presently employed, but a systematic comparison between these incubation systems with respect to preservation of slice function has not been performed yet. Therefore, we started a comparative study to evaluate five of these systems: the shaken flask (an Erlenmeyer in a shaking water bath), the stirred-well (24-well culture plate equipped with grids and magnetic stirrers), rocker platform (6-well culture plate with Netwell insert rocked on a platform), the roller system (dynamic organ culture rolled on an insert in a glass vial), and the 6-well shaker (6-well culture plate in a shaking water bath). The liver slices were incubated in these incubation systems for 0.5, 1.5, and 24.5 h and subsequently subjected to viability and metabolic function tests. The viability of the incubated liver slices was evaluated by: potassium content, MTT assay, energy charge, histomorphology, and LDH leakage. Their metabolic functions were studied by determination of the metabolism of lidocaine, testosterone, and antipyrine. Up to 1.5 h of incubation all five incubation systems gave similar results with respect to viability and metabolic function of the liver slices. However, after 24 h, the shaken flask, the rocker platform, and the 6-well shaker incubation systems appeared to be superior to the stirred well and the roller incubation systems.

Animals↗
PubMed2002

Influence of long-term changes in incubation temperature on catecholamine levels in plasma of chicken embryos (Gallus gallus f. domestica).

Catecholamine concentrations were determined from day 18 to 21 of incubation (D18, D21) in developing chicken embryos. The control group was continuously incubated at 37.5 degrees C. The eggs of the two other groups were incubated at 37.5 degrees C until day 14. In the cold group, temperature was decreased to 35.0 degrees C and in the warm group, incubation temperature was increased to 38.5 degrees C for the remainder of incubation. Plasma catecholamine concentrations were measured in eggs exposed to a change in incubation temperature for 4, 5, 6 and 7 days. Embryos in the warm group had dopamine (DA) and noradrenaline (NA) concentrations that were significantly higher than in the control group. On the contrary, eggs incubated at the cooler temperature had hormone levels that were significantly lower than in the control group. Adrenaline (A) levels in the two experimental treatments were significantly lower compared to control eggs. Temperature modulated the time needed for development. Chicken embryos are supposed to hatch on day 21. However, on day 20, NA concentration in the cold-incubated group was too low to fulfill its essential physiological function, whereas in the warm group, the NA concentration seems to be sufficient. Long-term exposure to altered incubation temperature affects the quantitative catecholamine concentration during development, but the relative proportion of each catecholamine remained constant.

Animals↗
PubMed2004

Effect of lighted incubation on embryonic growth and hatchability performance of two strains of layer breeder eggs.

1. Eggs from two layer-type breeder flocks (ISA-W vs Leghorn) between 30 and 45 weeks of age were used in 4 trials to study the effects of lighted incubation on embryonic growth from 5 to 18 d of age and hatchability per cent (HP). The physical dimensions, eggshell characteristics and conductance (EC) of eggs of the two strains were compared. 2. Eggs were set in an incubator on trays either in the dark-control or under two tubes of 20-watt white fluorescent light during the first 18 d of incubation. The light intensity ranged from 1230 to 1790 lux at the surface of the eggs. Eggs were transferred to dark hatching compartments at d 19 of incubation. 3. The genetic make-up of birds influenced the physical dimensions and eggshell characteristics of eggs. ISA-W eggs had higher weight, surface area (ESA), volume (EV), width, EC, shell volume and HP, and lower ESA:EV ratio, per cent shell, shell density and dead embryos than those of Leghorn eggs. 4. Lighted incubation increased daily embryonic growth (mg/d) and HP by 3.9 and 5.9%, respectively, when compared with the dark-control incubation. 5. The physical dimensions and eggshell characteristics of eggs influenced the effects of lighted incubation on embryonic growth and HP. Lighted incubation increased embryonic growth and HP of ISA-W eggs. The increase in embryonic growth and HP of the Leghorn eggs was not significant. 6. The genetic make-up of birds influenced the physical dimensions and eggshell characteristics of their eggs and these differences in the characteristics of hatching eggs influenced embryonic growth and HP when incubated under light.

Animals↗
PubMed2006

Effect of electric field during incubation of eggs on the immune responses of hatched chickens.

The effects of electric field (EF) during incubation of eggs on the immunocompetence of chickens were investigated over a 42-day experimental period. Eggs from a meat-type breeder flock were incubated under EF of 30 kV/m, 60 Hz during the first 18 days of incubation as compared with the control incubation (C). Chickens from the two incubation treatments were fed ad libitum and their immune system were monitored. Measurements were made of body weight (BW), and lymphoid organs weight (thymus, spleen, and bursa of Fabricius (BOF)) of birds at 21 and 42 days of age. Immune systems of birds were tested for specific antibody responses to sheep red blood cell (SRBC) and Newcastle disease vaccine (NDV), in vivo T-lymphocyte proliferation responses to phytohemagglutinin (PHA), and in vitro to concanavalin A (Con-A). EF incubation of eggs did not significantly (P > 0.05) influence BW of bird, absolute weight of lymphoid organs and weight of thymus, and BOF as a percentage of BW of bird (% BW) at 21 and 42 days of age, humoral immune responses as measured by antibody responses to SRBC and NDV, and cell-mediated immune responses as measured by T-lymphocyte proliferation responses to PHA, and Con-A of birds when compared with those of the C treatment. EF incubation of eggs significantly (P < 0.05) increased spleen weight as a % BW at 21 and 42 days of age when compared with those incubated under the C treatment. Birds at 42 days of age had significantly (P < 0.01) higher BW, lymphoid organ weight, and weight of BOF as a % BW, and lower spleen weight as a % BW when compared with those of 21 days of age. It is concluded that the incubation of eggs under EF of 30 kV/m, 60 Hz increased spleen weight as a % BW, without altering cell-mediated and humoral immune responses and, consequently, immunocompetence of meat chickens during the rearing period of 42 days.

Animals↗
PubMed2005

A prospective study, using sibling oocytes, examining the effect of 30 seconds versus 90 minutes gamete co-incubation in IVF.

BACKGROUND: Traditionally oocytes have been exposed to sperm overnight, for 16-20 h. This long period of co-incubation, however, has been shown to create problems with high levels of reactive oxygen species (ROS), which may affect embryo viability and cause hardening of the zona pellucida. Recently, a positive effect of reducing the co-incubation time to 90-120 min was reported. The objective of this study was to evaluate whether a further reduction of the co-incubation period could benefit the outcome of IVF. METHODS: In this prospective study, 777 sibling oocytes from 81 women undergoing IVF were divided via alternate allocation to co-incubation for either 30 s (ultrashort co-incubation) (group A) or for 90 min (standard co-incubation) (group B). Endpoints were normal fertilization (two-pronuclear, 2PN), polyspermy (>2PN), embryo quality (EQ), clinical pregnancy (CP) and implantation (IR). RESULTS: The normal fertilization rates of the two groups were comparable: group A 58.6% versus group B 58.0%. Significantly lower rates of polyspermy were seen in group A compared to group B (2.8 versus 7.2%, P = 0.008). No statistically significant differences in EQ, CP or IR were seen. CONCLUSION: This is the first study demonstrating the achievement of good fertilization rates in IVF with ultrashort co-incubation. Significantly lower rates of polyspermy were seen in the group with ultrashort compared to the standard co-incubation group. Further studies are, however, needed in order to evaluate whether ultrashort co-incubation has any effect on the outcome of IVF.

Adult↗
PubMed1997

Effect of strain and age of the broiler breeder female on incubation time and chick weight.

Two experiments were conducted to evaluate effects of strain [five in Experiment (Exp.) 1 and six in Exp. 2)] and age (29, 47, and 57 wk in Exp. 1 and 29, 41, and 52 wk in Exp. 2) of commercial broiler breeders on incubation time and chick weight. Highly significant differences in egg weight were found among strains in both Exp. After adjusting for effects of egg weight, significant effects of strain, age, and their interactions were found on incubation time, egg weight at transfer, and chick weight at hatch in Exp. 1, but not in Exp. 2. Mean incubation times varied among strains from 496.6 to 498.8 h in Exp. 1 and from 499.3 to 501.9 h in the second experiment. In Exp. 1, incubation time decreased from 498.6 h when breeders were 29 wk to 494.8 at 47 wk, whereas in Exp. 2, it decreased from 510.5 h at 29 wk to 495.1 h at 41 wk. This decrease also resulted in a negative correlation between egg weight and incubation time. Differences due to strain and age were found for yolk and albumen percentage and yolk: albumen ratio. Percentage yolk was 27.2 and 32.7% and percentage albumen was 60.1 and 55.9% in eggs from 29 to 52 wk breeders, respectively. Shell percentage was significantly affected by strain. Strain by age interactions were found for each response in Exp. 1 but only for set and chick weight in Exp. 2. Differences among incubators were found only for incubation time; interactions of incubation time and strain and age were also detected. Results indicate that genotype, age of the female breeder, and incubator should be considered along with their interactions to obtain optimum hatching performance.

Aging↗
PubMed2006

The effects of suboptimal eggshell temperature during incubation on broiler chick quality, live performance, and further processing yield.

Different incubation conditions can cause eggshell temperature (EST) to deviate from optimum. Two experiments were performed to determine the effect of low EST at the start of incubation and high EST at the end of incubation on hatchability, chick quality, 6-wk live performance, and breast meat yield of broiler chickens. In each experiment, 1,800 eggs from a single flock were divided and set into 2 setters. From 0 to 10 d of incubation, one setter was set to attain an EST of 36.6 degrees C (considered low), whereas the other was set to 37.8 degrees C (the control temperature). Using an infrared thermometer, EST was measured daily on a sample of eggs to ensure treatment intentions. On d 11 of incubation, the temperature of the low EST setter was increased to 37.8 degrees C in synchrony with the other setter until transfer. On d 18 of incubation, eggs from both setters were combined into 2 equal groups and transferred to hatchers. The EST in one hatcher was set to 37.8 degrees C (control) and in the other to 39.5 degrees C (considered high) until 21 d of incubation. Hatched males were placed in battery cages (Experiment 1) or floor pens (Experiment 2) and reared on common feeds to 1 or 6 wk of age, respectively. Low EST in the first 10 d of incubation reduced hatchability, increased BW and chick yield, and reduced 1-wk gain compared with the control EST. Throughout rearing, BW was reduced for low EST chicks compared with control EST chicks; consequently, carcass, fillet, and tender weights were also reduced. High EST in the hatcher increased hatchability, and reduced BW, chick yield, and 1-wk gain compared with control EST in the hatcher. By 3 wk of age, there was no difference in BW between chicks in high EST and control EST treatments. Subsequent carcass and processing yields were also similar. Incubation at the control EST of 37.8 degrees C, particularly from 0 to 10 d, resulted in the best performance overall.

Animals↗
PubMed1991

Ovarian steroid production in vitro during gonadal regression in the turkey. I. Changes associated with incubation behavior.

The mechanism regulating ovarian regression during incubation behavior in the domestic turkey has not been elucidated. This study was designed to determine whether ovarian steroidogenic potential is depressed during gonadal regression associated with the onset of incubation behavior. Hens were housed in floor pens equipped with trap nests that were checked 7 times per day. Hens were grouped, according to nesting frequency and egg production, into the following classifications: laying (laid an egg every day and trapped in the nest only once/day); transitional (laid an egg every day but trapped in the nest 4 or more times/day); and Day 1, Day 3, and Day 5 incubating (no egg for 2, 4, or 6 days, respectively, while trapped in the nest at least 4 times/day). Follicular atresia was evident in the largest preovulatory follicle (F1) in transitional hens, extensive in F1 through the third largest follicle (F3) in Day 1 incubating hens, and extensive in F1 through F7in Day 3 incubating hens. Levels of circulating LH, progesterone (P), androgen (A), and estradiol (E) decreased in transitional hens relative to concentrations in laying hens and remained low thereafter. In contrast, levels of prolactin were greater in Day 3 and Day 5 incubating hens than in laying, transitional, or Day 1 incubating hens. Basal production of P by F1 granulosa cells was lower from Day 1 incubating hens than from the other groups. Production of P in response to porcine-luteinizing hormone (pLH) was greater by cells from transitional and Day 1 incubating hens than from those of laying hens.(ABSTRACT TRUNCATED AT 250 WORDS)

Androgens↗
PubMed1996

Significance of pre-incubation temperature and inoculum concentration on subsequent growth of Listeria monocytogenes at 14 degrees C.

The influence of the bacterial concentration of an inoculum (10(1) or 10(3) cfu ml-1) of two strains of Listeria monocytogenes (Scott A: serotype 4b and V7: serotype 1) and one strain of L. innocua (Lin 11), and the time and temperature at which the inoculum was stored (cold storage: 4 degrees C for 4 weeks, or without cold storage: -20 degrees C before immediate transfer), and the temperature at which cells were pre-incubated (30 degrees C and 14 degrees C) on subsequent growth in Richard's broth at 14 degrees C was investigated. Richard's broth at a pH 5.9 was used to simulate potential growth in soft cheese (camembert type) and an incubation temperature of 14 degrees C was used to simulate storage-temperature ripening of cheese. Enumeration of the number of viable cells was by plate count method, except where viable cell numbers were less that 10(3) cfu ml-1, when the MPN (Most Probable Number) technique was used. With cold storage and an inoculum of 10(3) cfu ml-1 (high bacterial concentration) the pre-incubation temperatures (30 degrees C and 14 degrees C) did not significantly influence the subsequent growth curve: there was no significant lag (less that 21 h) and cell numbers peaked in about 8.5 d. However, with cold storage and an inoculum of 10(1) cfu ml-1 (low bacterial concentration) and a pre-incubation temperature of 30 degrees C a significant shift in the growth curve was observed over that pre-incubated at f14 degrees C, with the appearance of a lag of about 7.7 d. At a pre-incubation temperature of 14 degrees C with the low inoculum concentration, there was a measurable lag of about 1 d. Without cold storage and a pre-incubation temperature of 30 degrees C, there was a lag time of 2.3 d. Storage conditions, pre-incubation temperature and inoculum concentration therefore appear to influence the subsequent growth curve. Importantly, however, the growth curves for cultures from inocula, pre-incubated at either 30 degrees C or 14 degrees C, appeared to involve two distinct values of the exponential growth rate (k): the initial portion of the growth curve described by a low value of k and the subsequent portion by a consistently and significantly greater value. The appearance of two distinct growth phases was reproduced in further data determined for all the studied strains of the microorganism. Further study to explain these unexpected and reproducible findings is being conducted.

Colony Count, Microbial↗
PubMed2005

Influence of epithelium and isoprenaline incubation on responsiveness of guinea-pig trachea to methacholine.

There are reports regarding harmful effect of long-term use of beta2-agonist drugs on asthma severity and airway responsiveness. In the present study, the responses of guinea pig trachea with intact and denuded epithelium (groups 1 and 2, n = 10) to methacholine as EC50 were measured in tissues nonincubated or incubated with 10 mumol/l isoprenaline during the resting period. The same protocol was performed in groups 3 and 4 (n = 5 for each group) with an additional 30 min rest time after isoprenaline incubation. The response of trachea with denuded epithelium (groups 2 and 4) to methacholine was significantly higher than that with intact epithelium both in incubated and nonincubated conditions (groups 1 and 3, p < 0.05 to p < 0.001). Incubation with isoprenaline caused a significant reduction in the tracheal response to methacholine in both the denuded groups (p < 0.005 and p < 0.001) and intact epithelium groups (p < 0.005 for both cases). The reduction in tracheal responsiveness to methacholine due to incubation in epithelium denuded trachea (groups 2 and 4) was nonsignificantly greater than that of intact epithelium tissues. There was no difference between groups 3 and 4 with those of groups 1 and 2 in both incubated and non incubated conditions. The maximum contractility response to methacholine was not different between tracheal chains with denuded and intact epithelium and did not change due to incubation with isoprenaline. The results of this study indicate reduction of tracheal response to methacholine due to incubation of tissues with isoprenaline, which was relatively more pronounced in epithelium denuded trachea.

Adrenergic beta-Agonists↗
PubMed1979

Increases in mitochondrial steroidogenesis after short term incubation of porcine ovarian follicles with luteinizing hormone.

[4-14C]Cholesterol side chain cleavage, progesterone synthesis, and cytochrome oxidase activities were measured in mitochondria from unincubated and short term incubated large (8-10 mm) follicles isolated from porcine ovaries. Compared to the activity of mitochondria from unincubated follicles, specific [4-14C]cholesterol side chain cleavage activity in mitochondria from follicles incubated with LH (0.05 microgrogram/ml) did not change significantly after 12 h, but increased almost 2-fold after 18 h and 5-fold after 24 h. Also, specific mitochondrial progesterone synthesis activity increased dramatically after incubation of follicles for 24 h with LH. In comparison, mitochondria prepared from follicles incubated without LH showed no significant change in specific [4-14C]cholesterol side chain cleavage or progesterone synthesis activities after 18 h of incubation. While both of these activities increased after incubating follicles 24 h without LH, the values were significantly lower than those observed for preparations from follicles incubated with LH. In contrast to these changes in mitochondrial steroidogenesis, specific cytochrome oxidase activity in mitochondria did not change after incubation of follicles without or with LH. It is concluded that incubation of follicles with LH stimulates the development of mitochondrial steroidogenesis but initially does not affect some components of the respiratory chain.

Animals↗
PubMed1985

Behavioural and physiological effects of prolactin in incubating ring doves.

The role of prolactin in the maintenance of incubation behaviour in ring doves was re-examined and the dose-response relationships for behavioural, target tissue and body weight changes induced by injections of prolactin were compared in doves tested during the incubation phase of the breeding cycle. Doves given injections of prolactin twice a day starting on day 4 of incubation, during a 10-day period of isolation from their mates and nests, showed a higher persistence of incubation behaviour than doves injected with saline vehicle. However, the prolactin treatment failed to maintain incubation behaviour to the same extent as that observed in non-isolated untreated breeding pairs. Liver and body weights were higher and testicular weights lower in birds treated with high doses of prolactin than in non-isolated birds which had been incubating for 14 days. Good dose-response relationships were established between body, liver, crop and testes weights and the dose of prolactin administered. However, only a weak dose-response relationship was observed between prolactin and the maintenance of incubation behaviour. Overall, females injected with prolactin displayed more quiet sitting behaviour, less body weight gain and more gonadal regression than males injected with prolactin. Males in untreated breeding pairs had higher liver weights and lower crop weights than females. It is concluded that prolactin plays a role in maintaining readiness to incubate in doves, but that other factors may also contribute to this response. Further, it appears that prolactin mediates several target tissue changes which are sex-specific during incubation.

Animals↗
PubMedDate not supplied

Influence of incubation time/temperature on acrosome reaction/sperm penetration assay.

The acrosomal status of human spermatozoa was studied in relation to the score of the sperm penetration assay (SPA) at low-temperature (4 degrees C) incubation for induction and synchronization of the acrosome reaction (AR) and the incubation time of spermatozoa in conventional SPA. Spermatozoa were collected from 18 patients, selected by the "swim-up" method and treated in three different ways: (1) short-term incubation group (SIG): 3 h incubation at 37 degrees C, and (2) long-term incubation group (LIG): 20 h incubation at 37 degrees C, and (3) low temperature group (LTG): 24 h incubation at 4 degrees C followed by additional incubation at 37 degrees C for 3 h. The conventional methods of incubation, i.e. SIG (3 h) and LIG (20 h) did not show any significant differences as evaluated by the sperm penetration rate and the number of decondensing sperm heads per oocyte. In contrast, in the LTG all parameters were significantly increased, especially those of penetration rate (p less than 0.0005) and decondensing sperm heads per oocyte (p less than 0.0005). The percentage of AR significantly increased (p less than 0.0005) in the LTG (14.7%) compared with SIG (6.1%) and LIG (10.6%). A significant correlation was demonstrated between AR and the parameters used for evaluation of the SPA. The penetration rate (Spearman test, r = 0.462, n = 54, p less than 0.003) was the most significant parameter correlated with AR. It would appear that only human spermatozoa having completed AR are capable of fusing with the zona-free hamster ova.(ABSTRACT TRUNCATED AT 250 WORDS)

Acrosome↗
PubMed1993

Factors affecting insulin responsiveness of triglyceride synthesis in incubated chicken hepatocytes.

A technique was developed to isolate and incubate hepatocytes from 4-day-old chickens. Hepatocytes incubated in serum-free medium containing 1 ng chicken insulin/ml had greater rates of triglyceride (TG) synthesis than insulin-unexposed cells. Greater concentrations of insulin enhanced TG synthesis. Addition to medium of a supplement containing several hormones greatly improved insulin responsiveness of TG synthesis in hepatocytes compared with cells not incubated in supplemented medium. The medium supplement did not affect TG synthesis in hepatocytes incubated in the absence of insulin. The presence, compared with the absence, of triiodothyronine in the medium supplement improved insulin responsiveness of TG synthesis. In hepatocytes incubated in supplemented medium, increasing chicken insulin concentrations augmented malic enzyme activity. Hepatocytes incubated in supplemented medium containing bovine insulin had lesser rates of TG synthesis than cells incubated with similar concentrations of chicken insulin. To determine whether insulin responsiveness of TG synthesis is affected by age of chicken, hepatocytes from 4-, 11-, and 74-day-old chickens were incubated in supplemented medium. Responsiveness of TG synthesis to chicken insulin progressively decreased in hepatocytes from older chickens. Results indicate that hepatocytes isolated from 4-day-old chickens and incubated in supplemented medium are responsive to dilute insulin concentrations and may be useful to investigate the biochemical effects of this hormone.

Acetates↗
PubMedDate not supplied

Networked incubators. Hothouses of the new economy.

Business incubators such as Hotbank, CMGI, and Idealab! are a booming industry. Offering office space, funding, and basic services to start-ups, these organizations have become the hottest way to nurture and grow fledgling businesses. But are incubators a fleeting phenomenon born of an overheated stock market, or are they an important and lasting way of creating value and wealth in the new economy? The authors argue that one type of incubator, called a networked incubator, represents a fundamentally new and enduring organizational model uniquely suited to growing businesses in the Internet economy. It shares certain features with other incubators--mainly, it fosters a spirit of entrepreneurship and offers economies of scale. But its key distinguishing feature is its ability to give start-ups preferential access to a network of potential partners. Such incubators institutionalize their networking--they have systems in place to encourage networking, helping start-ups, for example, to meet with potential business allies. That doesn't mean incubatees get preferential treatment; it means only that they have built-in access to partnerships that might not have existed without the incubator. Even with this advantage, however, networked incubators can easily follow the road to ruin. To avoid failure, they must create a portfolio of companies and advisers that their incubatees can leverage. That can be done by strategically investing in portfolio firms and by enlisting a large set of business allies. It can also be done by establishing connections and relationships that are anchored more to the incubator than to particular individuals.

Cooperative Behavior↗
PubMed1992

Effect of prolonged incubation of male rat whole pituitary or pituitary-hypothalamus complex with testosterone on release of gonadotrophin and prolactin in vitro.

Investigations were undertaken to study the effect of in vitro addition of testosterone (0.3 mM) on the release of luteinizing hormone (LH), follicle stimulating hormone (FSH) and prolactin (PRL) by pituitary-hypothalamus complex (PHC) or the whole pituitary (PI) incubated for 72 hr, with incubation media changed every 24 hr. PHC or PI were from adult intact or castrated (7 days post castration) rats. The tissues incubated with or without testosterone were further exposed to 0.1 nM luteinizing hormone-releasing hormone (LHRH) for 4 hr. Incubation media and the pituitary were analyzed for PRL and gonadotrophin content. While PHC from normal and castrated rats released increasing amounts of LH with diminishing amounts of FSH and PRL at different periods of incubation, PI showed a decrease in the amounts of gonadotrophin and PRL released. Co-incubation of PHC or PI of intact or castrated rats with testosterone stimulated the release of LH and FSH during the first or second-24 hr incubation but inhibited the release of PRL in all the three incubations of 24 hr each. The extent of PRL inhibition increased with increasing incubation period. Testosterone had no effect on LHRH induced release of PRL but inhibited LHRH induced release of LH and FSH by pituitaries from constructs of normal rats. Testosterone reduced intrapituitary contents of PRL and FSH of intact and castrated rats. The data are interpreted to suggest that hypothalamus is essential for the maintenance of functional pituitary in vitro and that intrinsic differences exist in mechanisms regulating the secretion of LH, FSH and PRL.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1991

ELISA incubation times can be reduced by 2.45-GHz microwaves.

The ELISA (Enzyme Linked Immuno Sorbent Assay) technique is widely applied in the field of immunology. The use of this technique implies several incubation periods, often requiring more than one hour each. Microwave irradiation is known to be very useful in accelerating different kinds of processes. This knowledge has led to the present study in which it is proved that 2.45-GHz microwaves are able to reduce ELISA incubation times. This time reduction of 50% or more concerns each of the four incubation steps of the specific indirect ELISA which is used to determine the amount of antibodies in NF90-hybridoma cell culture supernatants. These incubation steps involve the coating of the microtiter plate, the incubation with BSA against non-specific binding, and the incubations with the first and second antibody. Combinations of more microwave-influenced steps in one ELISA save a considerable amount of time. When in addition air is blown through the incubation fluid during microwaving, the extinction values are almost the same as those of conventionally performed ELISAs. Thus, a good match of power setting and irradiation time results in a total incubation time reduction from 5.5 hours to 2 hours in this particular ELISA. Carefully monitoring the temperature of the incubation fluids during irradiation with the use of a fiberoptic thermometer appeared to be of crucial importance in the development of an optimal procedure.

Enzyme-Linked Immunosorbent Assay↗
PubMed1985

Effect of atmosphere of incubation on the isolation of group A streptococci from throat cultures.

The optimal atmosphere of incubation for the isolation of group A beta-hemolytic streptococci from throat cultures has been the subject of considerable debate. To resolve this issue, we examined 5992 consecutive throat cultures performed at a private pediatric office in children with acute upper respiratory tract infections. All throat cultures were streaked onto duplicate blood agar plates, one of which was then incubated anaerobically and the other aerobically. beta-Hemolytic streptococci were isolated in cultures from 1885 (31.5%) of the patients; 1479 (24.7%) were identified as group A, and 406 (6.8%) were identified as non-group A. Group A streptococci were recovered significantly more often from the plates incubated anaerobically than from those incubated aerobically (1467 vs. 1054; anaerobic only, 425; aerobic only, 12; P less than 0.01). Non-group A streptococci were also recovered significantly more often from the plates incubated anaerobically than from the plates incubated aerobically (397 vs. 170; anaerobic only, 236; aerobic only, 9;P less than 0.01). Anaerobic incubation maximizes the yield of the throat culture. The additional cost and effort of anaerobic incubation are small, and would appear to be justified by the increased isolation rate of group A beta-hemolytic streptococci. The significance of the increased isolation rate of non-group A beta-hemolytic streptococci with anaerobic incubation needs to be investigated further.

Adolescent↗