For maceration of needle mesophyll samples, in order to count chloroplasts directly in separated cells, different methods were tested: a 1M aqueous solution of HCl (Possingham and Saurer, 1969); a 3.5% aqueous solution of glutaraldehyde and 0.1M aqueous solution of Na2EDTA (Boffey et al., 1979); and a solution of 10% aqueous CrO3 and 10% aqueous HNO3 (OâBrian and McCully, 1981). Based on the above-presented lines of evidence, we conclude that the presented optical disector method for chloroplast counting, using stacks of confocal microscopic images acquired from thick tissue sections accompanied by SUR sampling, is a very efficient method for estimating chloroplast number per mesophyll cell.
current-year needles were collected from crowns of 18-year-old trees planted on an experimental site of the Global Change Research Centre, Academy of Sciences, Czech Republic, the BÃlÃ½ KÅÃÅ¾ in MoravskoslezskÃ© Beskydy mountains, in 2004 (Urban et al., 2001).
The images of the spruce needle cross-sections were captured using a Leica SP2 AOBS confocal laser-scanning microscope (Leica Microsystems, Wetzlar, Germany) with Ar laser excitation of 488nm; phenolics autofluorescence was detected in the green channel (494â577nm) and autofluorescence of chlorophyll in chloroplasts was detected in the red channel (625â710nm) (Fig. 2007), but to our knowledge it has never been used for chloroplast number estimation.
Control of starch granule numbers in Arabidopsis chloroplasts. . M
Authors often do not describe the sampling design and the method used for chloroplast counting. J
In simpler terms, they are known as leaf cells.
One of the most frequently used methods for estimation of chloroplast number per mesophyll cell in herbaceous plants in 2D is counting chloroplasts in separated mesophyll cells obtained by maceration procedures, as described by Possingham and Saurer (1969). NS
We made 111 sections through the cell model and counted profiles in each section. Zellnig
CO2 concentration (Wang et al., 2004; Teng et al., 2006). Information and translations of palisade cell in the most comprehensive dictionary definitions resource on the web. CA
(C) Sampling frames were superimposed on a needle transverse section using the Rectangles module in Ellipse software.
. . The particles are sampled without bias if each particle has the same probability of being sampled, i.e. The particular mesophyll cell chosen for 3D reconstruction was a smaller one, so that it was possible to focus through the entire cell.
For an unbiased estimation of the number of particles in 3D with no assumption about particle shape and size, the optical disector method based on 3D unbiased sampling probe was developed (Sterio, 1984; Gundersen, 1986). SA
We determined that the systematic shift in an estimated value is inversely related to the ratio of the height of chloroplasts to the height of mesophyll cells. Frandsen
The spongy mesophyll is found towards the lower epidermis. M
Determining the chloroplast number per cell in 3D reconstructions made from a series of confocal microscope images (Mozafari et al., 1997; Dinkins et al., 2001; Coate et al., 2012; Xu et al., 2012) can yield an unbiased estimate if SUR sampling is applied and a sufficient number of cells is analysed. . T
. However, this is a much more time-consuming approach than application of the disector method. RM
Chloroplasts are cell organelles where photosynthesis takes place. C
The mean number of particle profiles per section was 17.23Â±0.69 (meanÂ±SE). Zechmann
4). Cell-type-specific transcription factors are key to deducing the distinct functions of specialized cells from gene expression profiles. YT
The chloroplast number per cell in conifers was estimated by profile counting to be six to 16 chloroplasts per cell in different species sampled in August (Maslova et al., 2009) (Supplementary Table S1). Lamppa
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