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Biomedical subjects

E Soini

Publications and source records attributed to E Soini.

At least 19 recordsLinked to original sources
PubMed2004

Perceived role of fibre in a healthy diet among Finnish consumers.

OBJECTIVE: The aim was to study how Finnish consumers perceive the role of fibre in the diet, which foods are regarded as good sources of fibre and the relationship between the respondents' self-estimated fibre intake and their measured intake. METHODS: A semistructured interview was conducted with 125 volunteers, including a background information questionnaire and an easy-to-use self-administered paper-and-pencil form estimating fibre intake. RESULTS: According to the self-administered form about half of the respondents had adequate fibre intake. Among those who estimated their fibre intake as adequate/maybe adequate, only 61% belonged to the highest fibre intake group. Most of the respondents defined their diet as being healthy. The key elements for a healthy diet were 'vegetables', 'low in fat', 'fruit and berries' and 'variety'. Only 5% of the respondents mentioned fibre spontaneously here. However, fibre was considered important for health because of its effect on bowel function and general well-being. The recommended intake of fibre could not be described in nutritional terms, but respondents could identify relevant sources of fibre in the diet. CONCLUSION: Finnish consumers considered fibre important for health and could recognize the sources of fibre correctly although they did not mention it spontaneously as a part of a healthy diet.

Adult↗
PubMed2000

A new microvolume technique for bioaffinity assays using two-photon excitation.

Bioaffinity binding assays such as the immunoassay are widely used in life science research. In an immunoassay, specific antibodies are used to bind target molecules in the sample, and quantification of the binding reaction reveals the amount of the target molecules. Here we present a method to measure bioaffinity assays using the two-photon excitation of fluorescence. In this method, microparticles are used as solid phase in binding the target molecules. The degree of binding is then quantified from individual microparticles by use of two photon excitation of fluorescence. We demonstrated the effectiveness of the method using the human alpha-fetoprotein (AFP) immunoassay, which is used to detect fetal disorders. The sensitivity and dynamic range we obtained with this assay indicate that this method can provide a cost-effective and simple way to measure various biomolecules in solution for research and clinical applications.

Fetal Diseases↗
PubMed2000

Two-photon fluorescence excitation in detection of biomolecules.

Two-photon fluorescence excitation has been found to be a very powerful method for enhancing the sensitivity and resolution in far-field light microscopy. Two-photon fluorescence excitation also provides a substantially background-free detection on the single-molecule level. It allows direct monitoring of formation of labelled biomolecule complexes in solution. Two-photon excitation is created when, by focusing an intensive light source, the density of photons per unit volume and per unit time becomes high enough for two photons to be absorbed into the same chromophore. In this case, the absorbed energy is the sum of the energies of the two photons. In two-photon excitation, dye molecules are excited only when both photons are absorbed simultaneously. The probability of absorption of two photons is equal to the product of probability distributions of absorption of the single photons. The emission of two photons is thus a quadratic process with respect to illumination intensity. Thus in two-photon excitation, only the fluorescence that is formed in the clearly restricted three-dimensional vicinity of the focal point is excited. We have developed an assay concept that is able to distinguish optically between the signal emitted from a microparticle in the focal point of the laser beam, and the signal emitted from the surrounding free labelled reagent. Moreover, the free labels outside the focal volume do not contribute any significant signal. This means that the assay is separation-free. The method based on two-photon fluorescence excitation makes possible fast single-step and separation-free immunoassays, for example, for whole blood samples. Since the method allows a separation-free assay in very small volumes, the method is very useful for high-throughput screening assays. Consequently we believe that two-photon fluorescence excitation will make a remarkable impact as a research tool and a routine method in many fields of analysis.

Dose-Response Relationship, Drug↗
PubMed1999

Photon-burst analysis in two-photon fluorescence excitation flow cytometry.

We studied the use of a dramatically reduced testing zone in combination with two-photon excitation and photon-burst analysis in high-throughput rare-event detection simulation using a modified flow cytometer. Two-photon excitation measurements were performed with a mode-locked titanium:sapphire laser. Fluorescence emission was measured with a photon-counting avalanche photodiode. Measured signal was analysed offline by autocorrelation and burst detection methods. Test samples were composed of full blood and orange fluorescent polystyrene nanospheres mixed in full blood. Results show that two-photon fluorescence excitation and time-correlation analysis provide a good signal-to-noise ratio for rare-event particle detection in a turbid sample environment.

Algorithms↗
PubMed1998

A new design of the flow cuvette and optical set-up for the scanning flow cytometer.

We introduce a new design for the optical cuvette and a new optical lay-out for the Scanning Flow Cytometer (SFC) that permits measurement of the angular dependency of the scattered light from individual moving particles. The improved optical scheme of the SFC allows measurement of the angular scattering pattern of individual particles at polar angles from 10 degrees to 120 degrees with integration at azimuthal angles from 0 degrees to 360 degrees and with angular resolution of better than 0.5 degrees. The performance of the SFC is demonstrated using certified polystyrene particles as reference material The aim of this work is to develop a flow cytometer, which, by recording the entire light scattering pattern of individual biological particles, would provide more information about the particle structure than the ordinary wide angle, forward and side scattering concepts.

Flow Cytometry↗
PubMed1997

Computer analysis of living cells: movements of the chromatoid body in early spermatids compared with its ultrastructure in snap-frozen preparations.

Quantitative analyses of cytoplasmic and nuclear organelle movements in living interphase cells at defined stages of differentiation are few. By phase contrast videomicroscopy and digital imaging techniques, we have traced the path of the chromatoid body (CB) and analysed its rapidly changing positions in relation to the nuclear envelope, Golgi complex and nuclear pale chromatin areas in living early spermatids of the rat. The CB had intimate interactions with the nuclear envelope and moved both in parallel and perpendicular fashion in relation to it. It had successive short contacts with the Golgi complex and nuclear pale chromatin areas. It was also seen to scan between two pale chromatin areas and it had pinocytosis-like transient engulfments during interactions with the pale chromatin. In ultrastructural analysis of snap-frozen preparations, the CB had a large contact area with the nuclear envelope with several intermediate organelles that may be involved in nucleocytoplasmic material transport. It is evident that quantitative image analysis of living cells is a powerful guide for ultrastructural analyses. The snap-freezing technique gives new possibilities for studies of structures that are sensitive to conventional fixation procedures.

Animals↗
PubMed1994

Use of fluorescent europium chelates as labels in microscopy allows glutaraldehyde fixation and permanent mounting and leads to reduced autofluorescence and good long-term stability.

In the present study europium chelates were introduced as alternative fluorescent labels for microscopy and their effect on enhanced autofluorescence caused by the glutaraldehyde fixative was investigated. Glutaraldehyde fixation was used to stabilize the cells for a permanent mount after the immunocytochemical reaction. The europium signal in time-resolved fluorescence microscopy was shown to be free of autofluorescence when strong cross-linking fixation with glutaraldehyde was used and the signal-to-background ratio obtained was 2,400 or better. It was also shown that the europium signal was stable in daylight and at room temperature. Fluorescent europium chelate used in this experiment provides excellent contrast and long-term stability for the samples with glutaraldehyde fixation and permanent mounting.

Antibodies, Monoclonal↗
PubMed1992

Time-resolved fluorescence imaging of europium chelate label in immunohistochemistry and in situ hybridization.

Fluorescent lanthanide chelates with long decay times allow the suppression of the fast decaying autofluorescence in biological specimens. This property makes lanthanide chelates attractive as labels for fluorescence microscopy. As a consequence of the suppression of the background fluorescence the sensitivity can be increased. We modified a standard epifluorescence microscope for time-resolved fluorescence imaging by adding a pulsed light source and a chopper in the narrow aperture plane. A cooled CCD-camera was used for detection and the images were digitally processed. A fluorescent europium chelate was conjugated to antisera and to streptavidin. These conjugates were used for the localization of tumor associated antigen C242 in the malignant mucosa of human colon, for the localization of type II collagen mRNA in developing human cartilaginary growth plates, and for the detection of HPV type specific gene sequences in the squamous epithelium of human cervix. The specific slowly decaying fluorescence of the europium label could be effectively separated from the fast decaying background fluorescence. It was possible to use the europium label at the cell and tissue level and the autofluorescence was effectively suppressed in in situ hybridization and immunohistochemical reactions in both frozen and formaldehyde-fixed, wax-embedded specimens.

Alkaline Phosphatase↗
PubMed1990

Time-resolved fluorescence in biospecific assays.

Fluorescent lanthanide chelates and their detection using time-resolved fluorometry for in vitro as well as for microscopic bio-specific assays are reviewed, with reference to other fluorescent dyes and labelling materials. The minimum detectable dose of labelled components and the theoretical ultimate sensitivities of immunoassays and DNA-assays are discussed and compared with experimental results. It is concluded that the time-resolved fluorometry provides a high sensitivity labelling method in immunoassays, and a sensitivity comparable to phosphorus-32 in DNA hybridization assays. It is also concluded that lanthanide chelates are a potential alternative for organic fluorescent dyes in microscopy and that they are most useful in multiparameter assays. The combination of organic fluorescent dyes and lanthanide chelates provides the highest separation efficiency between two assay parameters.

DNA Probes↗
PubMed1985

A new design for a liquid scintillation counter for micro samples using a flat-bed geometry.

A new design for a liquid scintillation counter based on a flat-bed geometry is described. Micro-samples are dried or filtered onto transfer membranes or glass fibre filters in a 6 X 16 matrix, compatible with 96-well micro-titration plate filtration assays of labelled cells. A prototype counter without lead shielding had low background countrates (2-3 cpm for 3H) giving a figure of merit of 1325 (and 1292 for 14C). Only 5-15 ml of scintillant/96 samples are required and thus the volume of radioactive waste is low.

DNA↗
PubMed1983

Detection of hepatitis B surface antigen using time-resolved fluoroimmunoassay.

Conventional fluoroimmunoassay (FIA) methods based on various fluorescence principles have not achieved the sensitivity of radioimmunoassay (RIA) mainly because of problems of background fluorescence arising, for example, from the biological specimen. We now describe an immunoassay of hepatitis B surface antigen (HBsAg) based on time-resolved (TR) fluorescence using a lanthanide as label. The assay initiates the development of a new generation of immunoassays. The fluorescence intensity is measured after a selected delay time which almost completely eliminates background fluorescence, which has a fast decay time. The excitation is performed with a flashing light source. The molecules with a long fluorescent lifetime consist of chelates of rare earth metals (Eu, Tb, Sm, Dy). They absorb strongly the excitation radiation and transfer the energy to the chelated central atom which in turn produces an emission spectrum characteristic of the lanthanide used. A long Stokes' shift (greater than 270 nm) helps to reduce the background in the emission region of the chelate and thus optimizes measurement of the relevant fluorescence. The present TR-FIA uses 2-naphthoyltrifluoroacetone as chelating agent because it creates an intense fluorescence with the rare earth metals. Synergistic agents such as trioctylphosphineoxid further enhance the fluorescence of the chelate. Depending on the instrumentation used for measuring time-resolved fluorescence and the conditions used for chelate formation, lanthanides can be detected at 10(-12)-10(-14)M concentrations.

Fluorescent Antibody Technique↗
PubMed1983

Time-resolved fluoroimmunoassay of human choriogonadotropin.

We describe time-resolved fluoroimmunoassay for human choriogonadotropin involving monoclonal antibodies directed against the beta- and alpha-subunits. The latter antibody was labeled with europium, which was measured by counting for 1 s after the immunoreaction was completed. In the solid-phase sandwich assay, both a one-step and two-step procedure were used; the respective measuring ranges were 0.7-135 and 0.7-350 int. units/L, the latter covering a 500-fold dynamic range. The CV within the assay range was between 4 and 8%, depending on the dose. Cross reactivity with lutropin in the one- and two-step procedures was 1.6% and 1.0%, respectively.

Antibodies, Monoclonal↗
PubMed1982

Versatile semiautomated sample processor and gamma counter to increase radioimmunoassay efficiency.

We describe a semiautomated batch-assay system designed to increase the speed and ease of performing radioimmunoassays while maintaining good accuracy and precision. The main components are a programmable pipetting unit (sample processor) and a gamma counter capable of simultaneously counting radioactivity in 12 samples. The processor uses prearranged sets of as many as 24 disposable pipette tips, which eliminates carryover between samples, and features a horizontal shaker and magnetic stirrer; 50 different assay protocols can be selected. Batches of tubes are easily transferred from the sample processor to the incubator and centrifuge. The gamma counter is controlled by a microprocessor, which is also used for data processing. Because the system is based on discrete rather than continuous-flow analysis and because of the ease with which consecutive samples can be processed, more than one operation and several different assays can be accommodated simultaneously, which saves time considerably.

Estriol↗
PubMed1981

Quantitative assessment of steroid hormone binding sites by thaw-mount autoradiography.

A procedure for the quantitative assessment of nuclear receptors for steroid hormones--and other substances--in individual cells is presented. Thaw-mount autoradiography, a procedure developed earlier in our laboratory, is utilized. The silver grain yield (specific activity) is 16.6 disintegrations per silver as determined fo tritium in guinea pig uterine tissues. An integrated formula is presented and applied for 3H-estradiol, 3H-diethylstilbestrol, and 3H-aldosterone in sampled tissue. A comparison with data derived from the literature that are based on the homogenization of whole uteri and biochemical analysis shows comparable values wtih the autoradiographic data if the latter are pooled. The pooled ata indicated 12-14,00 molecules of 3H-estradiol per uterine nucleus, while subpopulations of target cells vary between 5,000 and 28,000 per nucleus.

Animals↗