species and types that work WELL for analysis
Species and types that work well for analysis:
Genus that have been used for this type of research before are:
Genus that have been used for this type of research before are:
species and types do not work well for analysis
Some limu species do not report δ15N accurately.
These include:
- calcified algae (such as genus Jania, Galaura, Halimeda, Neomeris, and Padina)
- nitrogen fixing algae (such as genus Dictyospheria)
- cyanophytes (such as genus Lyngbya and Symploca)
These include:
- calcified algae (such as genus Jania, Galaura, Halimeda, Neomeris, and Padina)
- nitrogen fixing algae (such as genus Dictyospheria)
- cyanophytes (such as genus Lyngbya and Symploca)
Calcified Algae
Nitrogen-fixers and Cyanophytes
Sampling steps
1. Collect
Collecting Seaweed for Stable Isotope Analysis Seaweed (limu) should be collected at low tide When collecting the seaweed: Seaweed/limu (often but not always) needs a firm foundation to grow from and will often be found on rocky basaltic lava area or areas with limestone (coral) foundation. Pinch with your nail above the seaweed’s connection to the rock. Some seaweed (although not all have holdfast or connection to the substrate) have holdfast which connects them to the rock.
2. Clean
*If you can get distilled water ($1.25 per gallon at Walmart) it's best but if not tap water is fine too!
1. Using fresh water (distilled if possible), rinse the limu and remove dirt/sand/all things (epiphytes) growing on it besides the limu itself.
2. Repeat the rinse three additional times.
3. Pat down limu with paper towel three times
4. With limu inside, gather paper towel around the limu and shake vigorously 10 times.
3. Dry
Dry Samples Separate all samples by location in sterile zip lock bags with labels showing your name, the location/ sample number and the date that the sample was taken. The following treatments can be used to dry the algae (Markel 2017 unpub.): Treatment 1: Conventional Oven at 60°C (standard method used for this research) algae are baked in oven until the weight of the sample no longer changes (Nitrate Elimination Co 2012). Treatment 2: Herbarium, algae are pressed between wax paper and watercolor paper, then pressed between newspaper for absorption, then pressed in-between cardboard, and then have pressure applied on top Treatment 3: Silica gel is currently used for molecular analysis to dry samples for analysis (Roeder et al. 2004). Treatment 4: Sea salt, as Instant Ocean, was used historically for preservation of foods, and has desiccant ability.
4. Grind (Can be done by you or Lab technician, see Prepare to Sample!)
Grinding What you need: Ethanol, lab wipes, sterile two-part hand grinder, forceps, scraper, sterile lab specimen container labeled with the same information as the zip lock bags, goggles, lab coat, latex gloves Put on all lab-wear. Organize the bags of dried samples in order of location. Sterilize hand grinder and all utensils before and after each sample with ethanol and lab wipes. Remove a single sample from the first zip lock bag using sterilized forceps and place it in the grinder. Grind the sample until it becomes a fine powder. Carefully pour the ground up sample into the matching labeled sample container and scrap all the access powder using the scraper. Sterilize the grinder and utensils completely and begin the process again using the next sample.
5. Weighing and Analyzing
Balance What you need: Lab balancer, lab wipes ethanol, lab coat, goggles, and latex gloves, samples in specimen containers, log journal, two sterile lab forceps, and small tin lab balancer capsules, numbered small containers for finished samples Before each sample, sterilize forceps and table area using ethanol and lab wipes. Using previous information (if available) place the needed amount of the first sample into a prepped capsule. If a known amount is not provided for your specimen use around 2mg (use balance to weigh) and fold each corner of the capsule completely shut around the specimen and push the sides against a solid, clean surface. Then, lightly tap the closed capsule on its side. If any powder comes out, begin the process again. If no powder escapes, place the capsule into a numbered slot in the container and note the sample number, date, and the slot number in a personal or lab log journal. Sterilize all utensils and table space before beginning again with the second sample and repeat until all samples are completed.
Collecting Seaweed for Stable Isotope Analysis Seaweed (limu) should be collected at low tide When collecting the seaweed: Seaweed/limu (often but not always) needs a firm foundation to grow from and will often be found on rocky basaltic lava area or areas with limestone (coral) foundation. Pinch with your nail above the seaweed’s connection to the rock. Some seaweed (although not all have holdfast or connection to the substrate) have holdfast which connects them to the rock.
2. Clean
*If you can get distilled water ($1.25 per gallon at Walmart) it's best but if not tap water is fine too!
1. Using fresh water (distilled if possible), rinse the limu and remove dirt/sand/all things (epiphytes) growing on it besides the limu itself.
2. Repeat the rinse three additional times.
3. Pat down limu with paper towel three times
4. With limu inside, gather paper towel around the limu and shake vigorously 10 times.
3. Dry
Dry Samples Separate all samples by location in sterile zip lock bags with labels showing your name, the location/ sample number and the date that the sample was taken. The following treatments can be used to dry the algae (Markel 2017 unpub.): Treatment 1: Conventional Oven at 60°C (standard method used for this research) algae are baked in oven until the weight of the sample no longer changes (Nitrate Elimination Co 2012). Treatment 2: Herbarium, algae are pressed between wax paper and watercolor paper, then pressed between newspaper for absorption, then pressed in-between cardboard, and then have pressure applied on top Treatment 3: Silica gel is currently used for molecular analysis to dry samples for analysis (Roeder et al. 2004). Treatment 4: Sea salt, as Instant Ocean, was used historically for preservation of foods, and has desiccant ability.
4. Grind (Can be done by you or Lab technician, see Prepare to Sample!)
Grinding What you need: Ethanol, lab wipes, sterile two-part hand grinder, forceps, scraper, sterile lab specimen container labeled with the same information as the zip lock bags, goggles, lab coat, latex gloves Put on all lab-wear. Organize the bags of dried samples in order of location. Sterilize hand grinder and all utensils before and after each sample with ethanol and lab wipes. Remove a single sample from the first zip lock bag using sterilized forceps and place it in the grinder. Grind the sample until it becomes a fine powder. Carefully pour the ground up sample into the matching labeled sample container and scrap all the access powder using the scraper. Sterilize the grinder and utensils completely and begin the process again using the next sample.
5. Weighing and Analyzing
Balance What you need: Lab balancer, lab wipes ethanol, lab coat, goggles, and latex gloves, samples in specimen containers, log journal, two sterile lab forceps, and small tin lab balancer capsules, numbered small containers for finished samples Before each sample, sterilize forceps and table area using ethanol and lab wipes. Using previous information (if available) place the needed amount of the first sample into a prepped capsule. If a known amount is not provided for your specimen use around 2mg (use balance to weigh) and fold each corner of the capsule completely shut around the specimen and push the sides against a solid, clean surface. Then, lightly tap the closed capsule on its side. If any powder comes out, begin the process again. If no powder escapes, place the capsule into a numbered slot in the container and note the sample number, date, and the slot number in a personal or lab log journal. Sterilize all utensils and table space before beginning again with the second sample and repeat until all samples are completed.