CN102192901A - Method for imaging pollen surface ornamentation based on laser scanning cofocal microscope - Google Patents
Method for imaging pollen surface ornamentation based on laser scanning cofocal microscope Download PDFInfo
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- CN102192901A CN102192901A CN2011100610437A CN201110061043A CN102192901A CN 102192901 A CN102192901 A CN 102192901A CN 2011100610437 A CN2011100610437 A CN 2011100610437A CN 201110061043 A CN201110061043 A CN 201110061043A CN 102192901 A CN102192901 A CN 102192901A
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Abstract
The invention provides a method for imaging pollen surface ornamentation based on a laser scanning cofocal microscope. The method is characterized by comprising the following steps of: marking plant pollen grains by using 0.5% acridine orange; scanning a pollen grain sample by using laser with the wavelength of 488nm from an argon ion laser of the laser scanning cofocal microscope; collecting the series optical section images of the pollen grains by using an XYZ scanning program, wherein a collecting signal wavelength is 520-600nm; and using the cofocal software to reconstruct a three-dimensional image for the collected series optical section images. The method is simple to operate and can be used for observing and researching the pollen surface ornamentations of various plants.
Description
Technical field
The invention provides a kind of pollen surface texturing formation method, belong to the microscopic study technical field based on laser scanning co-focusing microscope.
Background technology
Laser scanning co-focusing microscope is made of microscope optical system, LASER Light Source, scanister and computer operating system, a whole set of instrument is by computer control, and the operation between each parts is switched and all can be carried out easily and flexibly in the computer operation platform interface.Laser scanning co-focusing microscope adopts laser as light source, detecting pinhole before utilization is placed on the illumination pin hole behind the light source and is placed on detecting device realizes that point throws light on and point is surveyed, the light of launching by the illumination pin hole from the light of light source focuses on certain point of sample focal plane, this institute's emitted fluorescence is imaged on the detecting pinhole, and the emission light beyond this point is detected pin hole to be stopped.Illumination pin hole and detecting pinhole are to illuminated point or to be detected a little be conjugation, so to be detected a little be confocal point, and the plane that is detected a place is confocal plane.
Use Laser Scanning Confocal Microscope, can gather tissue or cell interior the fluorescence labeling signal, observe the morphological change and the inner microtexture of variation, tissues observed or the cell of the interior ion concentration of living cells.Simultaneously, can also carry out the semi-quantitative analysis of three-dimensionalreconstruction and fluorescence labeling intensity to the image of gathering; Fluorescence in situ hybridization, the assignment of genes gene mapping and original position PCR in real time product are analyzed.At present, laser scanning co-focusing microscope has become the important means of cell biology, Neuscience, pharmacology, histological anatomy research.
Exposore decorative pattern feature is as one of reference frame of weighing angiosperm evolution degree, and is significant to its accurate description.Ordinary optical microscope can not be used for the observation of plant pollen surface texturing, observe the pollen surface texturing and generally need adopt scanning electron microscope, but the scanning electron microscope specimen preparation is comparatively complicated, apparatus expensive.The present invention adopts laser scanning co-focusing microscope observation of plant pollen surface texturing, and method is easy; Set up the method for visible light as light source analysis plant pollen surface texturing.
Summary of the invention
The purpose of this invention is to provide a kind of plant pollen surface texturing formation method based on laser scanning co-focusing microscope, its technical scheme is: plant pollen is positioned over microscope slide, adopted 0.5% acridine orange labeled pollen grain 5 minutes; Adopt the laser scanning pollen granule sample of laser scanning co-focusing microscope Argon ion laser 488nm wavelength, and adopt the XYZ scanning sequence to gather pollen granule series optical section image; The acquired signal wavelength is 520～600nm; And further adopt the burnt software of copolymerization that the serial optical section image of gathering is carried out 3-D view reconstruct.This method is simple to operate, can observe the plant pollen surface texturing.
The present invention compared with prior art has following advantage:
1, easy and simple to handle, cost is low;
2, adopt laser scanning co-focusing microscope visible light source herborization pollen surface texturing, imaging is clear;
3, range of application is wider, can be used for the research of various plant pollen surface texturings.
Embodiment
Embodiment 1
Get capsule of weeping forsythia pollen and be positioned over microscope slide, the acridine orange labeled pollen grain of employing 0.5% 5 minutes; Adopt the laser scanning Flos Forsythiae powder sample of laser scanning co-focusing microscope (German Lycra company produce model Leica TCS SP 2) Argon ion laser 488nm wavelength, the acquired signal wavelength is 520～600nm; And adopting the XYZ scanning sequence to gather Flos Forsythiae powder series optical section image, Flos Forsythiae powder series optical section image is as shown in Figure 1.Further adopt the burnt software of copolymerization (Leica TCS SP 2) that the serial optical section image of gathering is carried out 3-D view reconstruct, pollen reconstructed image and surface texturing are as shown in Figure 2.According to capsule of weeping forsythia pollen three-dimensionalreconstruction image, can observe Flos Forsythiae powder surface texturing.
Embodiment 2
Get red prince's weigela florida pollen and be positioned over microscope slide, the acridine orange labeled pollen grain of employing 0.5% 5 minutes; Adopt the red prince's weigela florida of the laser scanning pollen granule sample of laser scanning co-focusing microscope (German Lycra company produce model Leica TCS SP 2) Argon ion laser 488nm wavelength, the acquired signal wavelength is 520～600nm; And adopting the XYZ scanning sequence to gather pollen granule series optical section image, pollen granule series optical section image is as shown in Figure 3.Further adopt the burnt software of copolymerization (Leica TCS SP 2) that the serial optical section image of gathering is carried out 3-D view reconstruct, pollen reconstructed image and surface texturing are as shown in Figure 4.According to pollen three-dimensionalreconstruction image, can observe red prince's weigela florida pollen surface texturing.
Embodiment 3
Extracting honeysuckle pollen is positioned over microscope slide, the acridine orange labeled pollen grain of employing 0.5% 5 minutes; Adopt the laser scanning honeysuckle pollen granule sample of laser scanning co-focusing microscope (German Lycra company produce model Leica TCS SP 2) Argon ion laser 488nm wavelength, the acquired signal wavelength is 520～600nm; And adopting the XYZ scanning sequence to gather pollen granule series optical section image, pollen granule series optical section image is as shown in Figure 5.Further adopt the burnt software of copolymerization (Leica TCS SP 2) that the serial optical section image of gathering is carried out 3-D view reconstruct, pollen reconstructed image and surface texturing are as shown in Figure 6.According to pollen three-dimensionalreconstruction image, can observe honeysuckle pollen surface texturing.
Description of drawings
Fig. 1 shows Flos Forsythiae powder series optical section image;
Fig. 2 shows Flos Forsythiae powder three-dimensionalreconstruction image and surface texturing;
Fig. 3 shows red prince's weigela florida pollen granule series optical section image;
Fig. 4 shows red prince's weigela florida pollen granule three-dimensionalreconstruction image and surface texturing;
Fig. 5 shows honeysuckle pollen granule series optical section image;
Fig. 6 shows honeysuckle pollen granule three-dimensionalreconstruction image and surface texturing.
Claims (4)
1. the pollen surface texturing formation method based on laser scanning co-focusing microscope is characterized in that: (1) employing 0.5% acridine orange labeled pollen grain 5 minutes; (2) laser scanning, the images acquired of employing laser scanning co-focusing microscope Argon ion laser 488nm wavelength; The acquired signal wavelength is 520～600nm.(2) adopt laser scanning co-focusing microscope XYZ scanning sequence to gather pollen granule series optical section image, and further adopt the burnt software of copolymerization that the serial optical section image of gathering is carried out 3-D view reconstruct.This method is simple to operate, can observe the plant pollen surface texturing.
2. the plant pollen surface texturing formation method based on laser scanning co-focusing microscope as claimed in claim 1 is characterized in that: adopt 0.5% acridine orange labeled pollen grain, the mark time is 5 minutes.
3. the plant pollen surface texturing formation method based on laser scanning co-focusing microscope as claimed in claim 1 is characterized in that: the laser scanning, the images acquired that adopt laser scanning co-focusing microscope Argon ion laser 488nm wavelength; The acquired signal wavelength is 520～600nm.
4. the plant pollen surface texturing formation method based on laser scanning co-focusing microscope as claimed in claim 1, it is characterized in that: adopt laser scanning co-focusing microscope XYZ scanning sequence to gather pollen granule series optical section image, and further adopt the burnt software of copolymerization that the serial optical section image of gathering is carried out 3-D view reconstruct.
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| Application Number | Priority Date | Filing Date | Title |
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| CN2011100610437A CN102192901A (en) | 2011-03-10 | 2011-03-10 | Method for imaging pollen surface ornamentation based on laser scanning cofocal microscope |
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| CN2011100610437A CN102192901A (en) | 2011-03-10 | 2011-03-10 | Method for imaging pollen surface ornamentation based on laser scanning cofocal microscope |
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| CN102192901A true CN102192901A (en) | 2011-09-21 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104568880A (en) * | 2014-12-30 | 2015-04-29 | 北京大学第三医院 | Method for analyzing cartilage tissue |
| RU2717539C1 (en) * | 2019-04-10 | 2020-03-23 | Федеральное государственное бюджетное учреждение науки Сибирский федеральный научный центр агробиотехнологий Российской академии наук (СФНЦА РАН) | Method for determining botanical origin of honey |
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| CN101241069A (en) * | 2008-03-11 | 2008-08-13 | 武汉大学 | Color dispersion -type multifunctional Hadamard transform microscopical imaging spectrometer |
| CN101398381A (en) * | 2008-11-12 | 2009-04-01 | 南京农业大学 | Fluorescence labeling method for pear pollen and pollen tube fibril framework |
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2011
- 2011-03-10 CN CN2011100610437A patent/CN102192901A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101241069A (en) * | 2008-03-11 | 2008-08-13 | 武汉大学 | Color dispersion -type multifunctional Hadamard transform microscopical imaging spectrometer |
| CN101398381A (en) * | 2008-11-12 | 2009-04-01 | 南京农业大学 | Fluorescence labeling method for pear pollen and pollen tube fibril framework |
Non-Patent Citations (3)
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| 《中国体视学与图像分析》 19971231 陈耀文等 激光扫描共聚焦显微镜三维重建花粉的组织结构 223-225 1-4 第2卷, 第4期 * |
| 刘传银: "基于阿达玛变换显微成像的花粉形态及DNA定量研究", 《郧阳师范高等专科学校学报》 * |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104568880A (en) * | 2014-12-30 | 2015-04-29 | 北京大学第三医院 | Method for analyzing cartilage tissue |
| CN104568880B (en) * | 2014-12-30 | 2017-07-21 | 北京大学第三医院 | A kind of method analyzed cartilaginous tissue |
| RU2717539C1 (en) * | 2019-04-10 | 2020-03-23 | Федеральное государственное бюджетное учреждение науки Сибирский федеральный научный центр агробиотехнологий Российской академии наук (СФНЦА РАН) | Method for determining botanical origin of honey |
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Application publication date: 20110921 |