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Robust signal detection in 3D fluorescence microscopy
Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Matematisk-datavetenskapliga sektionen, Centrum för bildanalys. Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Matematisk-datavetenskapliga sektionen, Institutionen för informationsteknologi, Datoriserad bildanalys.
Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Matematisk-datavetenskapliga sektionen, Centrum för bildanalys. Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Matematisk-datavetenskapliga sektionen, Institutionen för informationsteknologi, Datoriserad bildanalys.
Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Matematisk-datavetenskapliga sektionen, Centrum för bildanalys. Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Matematisk-datavetenskapliga sektionen, Institutionen för informationsteknologi, Datoriserad bildanalys.
2010 (Engelska)Ingår i: Cytometry. Part A, ISSN 1552-4922, Vol. 77A, nr 1, s. 86-96Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

Robust detection and localization of biomolecules inside cells is of great importance to better understand the functions related to them. Fluorescence microscopy and specific staining methods make biomolecules appear as point-like signals on image data, often acquired in 3D. Visual detection of such point-like signals can be time consuming and problematic if the 3D images are large, containing many, sometimes overlapping, signals. This sets a demand for robust automated methods for accurate detection of signals in 3D fluorescence microscopy. We propose a new 3D point-source signal detection method that is based on Fourier series. The method consists of two parts, a detector, which is a cosine filter to enhance the point-like signals, and a verifier, which is a sine filter to validate the result from the detector. Compared to conventional methods, our method shows better robustness to noise and good ability to resolve signals that are spatially close. Tests on image data show that the method has equivalent accuracy in signal detection in comparison to Visual detection by experts. The proposed method can be used as an efficient point-like signal detection tool for various types of biological 3D image data.

Ort, förlag, år, upplaga, sidor
2010. Vol. 77A, nr 1, s. 86-96
Nationell ämneskategori
Bioinformatik och systembiologi
Identifikatorer
URN: urn:nbn:se:uu:diva-98015DOI: 10.1002/cyto.a.20795ISI: 000273384700011OAI: oai:DiVA.org:uu-98015DiVA, id: diva2:173171
Tillgänglig från: 2009-02-05 Skapad: 2009-02-05 Senast uppdaterad: 2011-11-04Bibliografiskt granskad
Ingår i avhandling
1. Digital Image Analysis of Cells: Applications in 2D, 3D and Time
Öppna denna publikation i ny flik eller fönster >>Digital Image Analysis of Cells: Applications in 2D, 3D and Time
2009 (Engelska)Doktorsavhandling, sammanläggning (Övrigt vetenskapligt)
Abstract [en]

Light microscopes are essential research tools in biology and medicine. Cell and tissue staining methods have improved immensely over the years and microscopes are now equipped with digital image acquisition capabilities. The image data produced require development of specialized analysis methods. This thesis presents digital image analysis methods for cell image data in 2D, 3D and time sequences.

Stem cells have the capability to differentiate into specific cell types. The mechanism behind differentiation can be studied by tracking cells over time. This thesis presents a combined segmentation and tracking algorithm for time sequence images of neural stem cells.The method handles splitting and merging of cells and the results are similar to those achieved by manual tracking.

Methods for detecting and localizing signals from fluorescence stained biomolecules are essential when studying how they function and interact. A study of Smad proteins, that serve as transcription factors by forming complexes and enter the cell nucleus, is included in the thesis. Confocal microscopy images of cell nuclei are delineated using gradient information, and Smad complexes are localized using a novel method for 3D signal detection. Thus, the localization of Smad complexes in relation to the nuclear membrane can be analyzed. A detailed comparison between the proposed and previous methods for detection of point-source signals is presented, showing that the proposed method has better resolving power and is more robust to noise.

In this thesis, it is also shown how cell confluence can be measured by classification of wavelet based texture features. Monitoring cell confluence is valuable for optimization of cell culture parameters and cell harvest. The results obtained agree with visual observations and provide an efficient approach to monitor cell confluence and detect necrosis.

Quantitative measurements on cells are important in both cytology and histology. The color provided by Pap (Papanicolaou) staining increases the available image information. The thesis explores different color spaces of Pap smear images from thyroid nodules, with the aim of finding the representation that maximizes detection of malignancies using color information in addition to quantitative morphological parameters.

The presented methods provide useful tools for cell image analysis, but they can of course also be used for other image analysis applications.

Ort, förlag, år, upplaga, sidor
Uppsala: Universitetsbiblioteket, 2009. s. 57
Serie
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 596
Nyckelord
Digital image analysis, microscopy, fluorescent staining, watershed segmentation, sub-cellular localization, point-like signals, wavelets, cell confluence, cytology, color spaces.
Nationell ämneskategori
Datorseende och robotik (autonoma system)
Identifikatorer
urn:nbn:se:uu:diva-9541 (URN)978-91-554-7398-3 (ISBN)
Disputation
2009-02-27, Siegbansalen, Ångstrom Laboratory, Polackbacken, Uppsala, 10:15
Opponent
Handledare
Tillgänglig från: 2009-02-05 Skapad: 2009-02-05 Senast uppdaterad: 2018-01-13Bibliografiskt granskad
2. Methods for 2D and 3D Quantitative Microscopy of Biological Samples
Öppna denna publikation i ny flik eller fönster >>Methods for 2D and 3D Quantitative Microscopy of Biological Samples
2011 (Engelska)Doktorsavhandling, sammanläggning (Övrigt vetenskapligt)
Abstract [en]

New microscopy techniques are continuously developed, resulting in more rapid acquisition of large amounts of data. Manual analysis of such data is extremely time-consuming and many features are difficult to quantify without the aid of a computer. But with automated image analysis biologists can extract quantitative measurements and increases throughput significantly, which becomes particularly important in high-throughput screening (HTS). This thesis addresses automation of traditional analysis of cell data as well as automation of both image capture and analysis in zebrafish high-throughput screening. 

It is common in microscopy images to stain the nuclei in the cells, and to label the DNA and proteins in different ways. Padlock-probing and proximity ligation are highly specific detection methods that  produce point-like signals within the cells. Accurate signal detection and segmentation is often a key step in analysis of these types of images. Cells in a sample will always show some degree of variation in DNA and protein expression and to quantify these variations each cell has to be analyzed individually. This thesis presents development and evaluation of single cell analysis on a range of different types of image data. In addition, we present a novel method for signal detection in three dimensions. 

HTS systems often use a combination of microscopy and image analysis to analyze cell-based samples. However, many diseases and biological pathways can be better studied in whole animals, particularly those that involve organ systems and multi-cellular interactions. The zebrafish is a widely-used vertebrate model of human organ function and development. Our collaborators have developed a high-throughput platform for cellular-resolution in vivo chemical and genetic screens on zebrafish larvae. This thesis presents improvements to the system, including accurate positioning of the fish which incorporates methods for detecting regions of interest, making the system fully automatic. Furthermore, the thesis describes a novel high-throughput tomography system for screening live zebrafish in both fluorescence and bright field microscopy. This 3D imaging approach combined with automatic quantification of morphological changes enables previously intractable high-throughput screening of vertebrate model organisms.

Ort, förlag, år, upplaga, sidor
Uppsala: Acta Universitatis Upsaliensis, 2011. s. 70
Serie
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 856
Nyckelord
Image analysis, cytmetry, model organism, zebrafish, screening
Nationell ämneskategori
Medicinsk bildbehandling
Forskningsämne
Datoriserad bildbehandling
Identifikatorer
urn:nbn:se:uu:diva-159196 (URN)978-91-554-8167-4 (ISBN)
Disputation
2011-11-11, Room 2446, Polacksbacken, Lägerhyddsvägen 2, Uppsala, 10:15 (Engelska)
Opponent
Handledare
Tillgänglig från: 2011-10-21 Skapad: 2011-09-25 Senast uppdaterad: 2014-07-21Bibliografiskt granskad

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