INTRODUCTION
Botryococcus braunii (B. braunii) yield some kind of hydrocarbons of which main component is similar to heavy
oil fuel (Figure 1). B. braunii discharge hydrocarbons to outside of the cells, while the other algae stock oils inside of
them. Hence it is expected that the heavy oil fuel can be continuously acquired from B. braunii without cell disruption.
However, the acquirable amount of the produced hydrocarbon is not yet enough to meet the production cost. One of the
reasons of the problem is depressed cellular function due to the damage caused by the toxicity of alcohols or organic
solvents used for deoiling of B. braunii [1]. To understand the effects of those chemicals on B. braunii and to improve
the deoiling process, a continuous assay of individuals is one of the most desired approaches for floating
microorganism such as B. braunii.
On the other hand, we had developed a microfluidic device which has a function to sample a small amount of the
individual microorganisms and line them up in the channel [2]. After sampling of them, some reagents can be
injected into the channel.
In this research, small amount of the cells of B. braunii were sampled in the device. Then ethanol and DAPI were
injected into the device for exposing B. braunii. We concurrently observed the effects of ethanol toxicity to
individual cells of B. braunii by confirming the fluorescence of the DAPI.
MICROFLUIDIC DEVICE
The microfluidic device made of PDMS has a capability to sample few amount of microorganisms. The function
is realized by a specific channel structure named as vernier walls which are standing on the ceiling and on the bottom
of the channel [2]. The closest vernier walls make the channel narrow periodically as shown in Figure 2. Hence,
microorganisms of uniform size are captured between the closest vernier walls. Additionally, to avoid clogging, the
microfluidic device has a bypass channel which almost microorganisms are going through
INTRODUCTION
Botryococcus braunii (B. braunii) yield some kind of hydrocarbons of which main component is similar to heavy
oil fuel (Figure 1). B. braunii discharge hydrocarbons to outside of the cells, while the other algae stock oils inside of
them. Hence it is expected that the heavy oil fuel can be continuously acquired from B. braunii without cell disruption.
However, the acquirable amount of the produced hydrocarbon is not yet enough to meet the production cost. One of the
reasons of the problem is depressed cellular function due to the damage caused by the toxicity of alcohols or organic
solvents used for deoiling of B. braunii [1]. To understand the effects of those chemicals on B. braunii and to improve
the deoiling process, a continuous assay of individuals is one of the most desired approaches for floating
microorganism such as B. braunii.
On the other hand, we had developed a microfluidic device which has a function to sample a small amount of the
individual microorganisms and line them up in the channel [2]. After sampling of them, some reagents can be
injected into the channel.
In this research, small amount of the cells of B. braunii were sampled in the device. Then ethanol and DAPI were
injected into the device for exposing B. braunii. We concurrently observed the effects of ethanol toxicity to
individual cells of B. braunii by confirming the fluorescence of the DAPI.
MICROFLUIDIC DEVICE
The microfluidic device made of PDMS has a capability to sample few amount of microorganisms. The function
is realized by a specific channel structure named as vernier walls which are standing on the ceiling and on the bottom
of the channel [2]. The closest vernier walls make the channel narrow periodically as shown in Figure 2. Hence,
microorganisms of uniform size are captured between the closest vernier walls. Additionally, to avoid clogging, the
microfluidic device has a bypass channel which almost microorganisms are going through
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