Automated, in vitro harvest of somatic embryos

Abstract

This work has demonstrated the aseptic, automated harvest of somatic embryos from a bioreactor suspension culture. Machine vision, emulating the selection criteria of an experienced biologist, classified embryos as harvestable or non-harvestable as they flowed through a 3 mm glass conduit. Embryos classified as harvestable were separated in a sealed harvest chamber. The system harvested 60% of the embryos at a rate of 2.4 embryos/h and incorrectly harvested less than 1% of the non-harvest objects. The low harvest rate precludes the applicability of this technique to research and commercial tissue culture laboratories. The suspension feed-rate, culture population density and culture homogeneity were identified as the most important factors influencing embryo harvest rate. The performance of this technique on more densely populated cultures was projected using anticipated improvements in suspension feedrate. It was concluded that, under the conditions of this analysis, the harvester would be of limited value in a commercial propagation environment but could be beneficial to many research labs working with plant somatic embryos.

Enhancement of somatic embryogenesis of Ipomoea batatas in solid eultures and production of mature somatic embryos in liquid cultures for application to a bioreactor production system

“…The computer projected 32 radii at equiangular increments from the centroid of an imaged embryo. Using the radii lengths and their points of intersection with the embryo perimeter, nine features describing embryo shape were measured, as defined in Harrell et al (1994).…”

“…Some embryos were visibly torn and bruised, and were not considered harvestable. When the reactors were inoculated with cultures containing mature embryos (Harrell et al 1994), the embryos were often damaged.…”

Application of imaging tools to plant cell culture: Relationship between plant cell aggregation and flavonoid production

“…Imaging tools were used to characterize plant cell embryo cultures grown in petri dishes for automatic sorting (10,11), and to assess their potential for conversion into plants (22) based on their morphology (40,41). Computer imaging was also coupled with standard or fluorescence microscopy using an external sampling loop or ex situ imaging for monitoring of microbial (42), yeast (26,36), fungal (39), and mammalian cell cultures (37).…”

“…The mean aggregate thickness (2 = 0.5r~ u with a = minor semiaxis) represents the light path through an aggregate. X v obtained from (o) manual and (o) automatic segmentation and as a function of the number of images processed;(11) represents the 95% confidence interval on the manually segmented estimates.FIG. 6.…”

Bioengineering Aspects Of Bioreactor Application In Plant Propagation

“…[2,[5][6][7][8][9][10][11]. The bioreactor technologies are also studied on their characteristics [5,[12][13][14][15][16] and on propagation of several plant species including shoots and somatic embryos [17][18][19][20][21][22][23][24][25][26][27][28][29].…”