This disclosure relates to a class of compounds which are utilized as biological stains in fluorescent microscopy. In particular, the disclosure relates to a method for detecting and identifying various structures in a biological sample which comprises treating said biological sample with a compound of the present disclosure to form a complex which emits fluorescence when irradiated with incident light. This class of compounds has been shown to be useful in detecting a variety of structures such as, for example, viruses, bacteria, yeasts, fungi, reticulocytes and cells in biological samples.
This is a division of application Ser. No. 278,812, filed June 29, 1981, now abandoned, which was a continuation-in-part of application Ser. No. 142,321, filed Apr. 21, 1980, now abandoned.
A method of measuring a melting temperature of a nucleic acid, which comprises a step of monitoring the fluorescent intensity of a mixture of a sample and a probe which is labeled with a fluorescent intercalative dye and contains a base sequence complementary to a specific nucleic acid in the sample, while varying the temperature of the mixture.
A method for detecting live individuals comprises a step of reacting a sample with a probe having a nonnatural type nucleic acid and a label substance linked with the nucleic acid, which nonnatural type nucleic acid contains a base sequence complementary to a base sequence of a target nucleic acid in the live target individuals to be detected and which probe is able to be incorporated into the live target individuals; and detecting the probe incorporated into the live target individuals by the utilization of the label substance of the probe when the live target individuals are present in the sample. A probe has a nonnatural type nucleic acid and a label substance linked with the nucleic acid, which nonatural type nucleic acid contains a base sequence complementary to a base sequence of a target nucleic acid in a live target individuals to be detected, and the probe is able to be incorporated into the live target individuals.
The present invention discloses a device and a process for quantitation of Toxoplasma gondii and Treponema pallidum antibody titer in a biological sample by immunofluorescent photometric microscopy. The process comprises: (a) reacting a specimen of said sample with an immunofluorescent reagent in a mounting medium containing a protective agent in an amount sufficient to reduce fading of a fluorescent reaction product less than 25% of initial fluorescent intensity; (b) localizing the specimen under transmitted, visible light; (c) reducing the effect of counterstain intensity by filters in emission light path; (d) measuring the sample using a fast shutter; (e) calibrating the photometer used in said microscopy by a stable fluorophore; (f) recording intensity of fluorescence of said specimen compared to standard negative and positive controls; (g) reducing effect of polar staining by substracting the corrected intensity of corresponding dilution of the negative control from the sample reading; and (h) assigning a numerical endpoint for serum antibody levels against Toxoplasma gondii or Treponema pallidum.
Reticulocytes, RNA or DNA are stained with a dye for detection in a flow cytometer. The dye has the formula: ##STR1## Wherein X=O, S, se or C(CH.sub.3).sub.2 ; R.sub.1 =alkyl having from 1-6 carbons; R.sub.2 =alkyl having from 1-6 carbons; R.sub.3 =fused benzene, alkyl (having 1-6 carbons), methoxy or is absent; R.sub.4 =alkyl having 1-6 carbons, methoxy or is absent; and n=zero or an integer from 1-6.
A method for multi-parameter analysis of cells in a body fluid sample is described which makes use of a plurality of fluorescence measurements, comprising at least two nucleic acid dyes and at least one fluorescently labelled cell surface marker, and a plurality of light scattering measurements. A kit containing the nucleic acid dyes and cell surface marker also is described.
